Compositions containing tucaresol or its analogs

ABSTRACT

Disclosed herein are compositions including a compound of formula (I) for treating cancer. Some embodiments relate to methods of treating cancer by co-administering a compound of formula (I) and one or more immune checkpoint inhibitor to a subject in need thereof. Some embodiments relate to methods of treating cancer by co-administering a compound of formula (I) and plinabulin to a subject in need thereof. Some embodiments relate to methods of providing co-stimulation of T-cell activation against cancer by co-administering a compound of formula (I), one or more immune checkpoint inhibitor.

INCORPORATION BY REFERENCE TO ANY PRIORITY APPLICATIONS

This application claims the benefit of U.S. Provisional Application No.62/292,763, filed Feb. 8, 2016, the disclosure of which is incorporatedherein by reference in its entirety.

BACKGROUND Field

The present invention relates to the field of chemistry and medicine.More particularly, the present invention relates to tucaresol and itsanalogs, compositions containing tucaresol or its analogs, and their usein treatment.

Description of the Related Art

Human cancers harbor numerous genetic and epigenetic alterations,generating neoantigens potentially recognizable by the immune system(Sjoblom et al, 2006) The adaptive immune system, comprised of T and Blymphocytes, has powerful anti-cancer potential, with a broad capacityand exquisite specificity to respond to diverse tumor antigens.

Recent cancer immunotherapy research has focused substantial effort onapproaches that enhance anti-tumor immunity by adoptive-transfer ofactivated effector cells, immunization against relevant antigens,providing non-specific immune-stimulatory agents such as cytokines, orremoving inhibitors to anti-cancer effector cells. Efforts to developimmune modulator and specific immune checkpoint inhibitors have begun toprovide new immunotherapeutic approaches for treating cancer, such asthe development of an antibody, ipilimumab, that binds to and inhibitsCytotoxic T-Lymphocyte Antigen-4 (CTLA-4) for the treatment of patientswith advanced melanoma (Hodi et al., 2010). In addition, nivolumab andpembrazumab, which are anti-PD-1 antibodies, have been approved fortreating melanoma, NSCLC and renal cancer, but patients only had limitedresponse to these therapies. While cancer remains as an incurabledisease for the great majority of patients, there exists a particularneed for developing effective therapeutic agents that can be used incancer immunotherapy.

SUMMARY OF THE INVENTION

Some embodiments relate to a pharmaceutical composition, comprising acompound of Formula (I):

and one or more immune checkpoint inhibitor.

Some embodiments relate to a pharmaceutical composition comprising acompound of Formula (I) and plinabulin.

Some embodiments relate to a method for treating a cancer, comprisingco-administering a compound of Formula (I) and one or more immunecheckpoint inhibitor to a subject in need thereof.

Some embodiments relate to a method for treating a cancer, comprisingco-administering a compound of Formula (I), one or more immunecheckpoint inhibitor; and plinabulin to a subject in need thereof.

Some embodiments relate to a method for treating a cancer, comprisingco-administering a compound of Formula (I) and plinabulin to a subjectin need thereof.

In some embodiments, the compound of Formula (I) described herein istucaresol.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 shows the tumor growth in each of the four treatment groupsincluding PD-1 antibody, PD-1 antibody/tucaresol, PD-1 and CTLA-4antibodies, and PD-1 antibody/CTLA-4 antibody/tucaresol.

FIG. 2 shows the anti-tumor effect of each of the four treatment groupsPD-1 antibody, PD-1 antibody/tucaresol, PD-1 and CTLA-4 antibodies, andPD-1 antibody/CTLA-4 antibody/tucaresol.

FIG. 3 shows the effect of tucaresol in combination with the PD-1antibody on MC38 tumor weight at necropsy.

FIG. 4 shows the MC 38 tumor growth in each of three treatment groupsincluding PD-1/CTLA-4 antibodies and PD-1 antibody/CTLA-4antibody/tucaresol.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENT

In some embodiments, compounds for use as described herein arerepresented by a compound of Formula (I),

-   -   Y¹ is selected from hydroxyl, C₁₋₄ alkylamino and acylamino        having a C₁₋₄ alkyl moiety thereof;    -   Y², Y³ and Y⁴ are independently selected from hydrogen, halogen,        C₁₋₄ alkyl, C₁₋₄alkoxy, trifluoromethyl, hydroxyl and benzyloxy;        and    -   Q¹ is either

where Q² and Q³ are independently selected from hydrogen and C₁₋₄ alkyl;

-   -   X is selected from cyano, carboxyl or a derivative thereof,        5-tetrazolyl and alkylsulfonylcarbamyl having a C₁₋₆ alkyl        moiety thereof; and    -   n is 0 or an integer selected from 1, 2, 3, 4, 5 and 6, and a        pharmaceutically acceptable salt thereof.

In some embodiments, when Y¹ is hydroxyl, Y², Y³ and Y⁴ are all

-   hydrogen and Q¹ is either then-   X is alkylsulfonylcarbamyl.

In some embodiments, the halogen for Y², Y³ and Y⁴ may be selected fromiodine, bromine, chlorine and fluorine.

In some embodiments, the alkyl for Q² and Q³ can independently have 1 to2 carbon atoms (i.e. methyl or ethyl). In some embodiments, the alkylfor Q² and Q³ can be methyl.

In some embodiments, X as a carboxyl derivative includes: esters,including aliphatic and aromatic hydrocarbon esters such as alkyl andaralkyl esters where for example the alkyl is C₁₋₁₂ alkyl and preferablyC₁₋₄ alkyl (in particular methyl, ethyl, isopropyl and t-butyl) andwhere the aralkyl is for example benzyl; and amides, including theunsubstituted amide, N-substituted amides and N,N-disubstituted amides(embracing cyclic and heterocyclic amides) where the substituentgroup(s) is (are) for example aliphatic hydrocarbon such as alkyl, inparticular C₁₋₆ alkyl such as methyl, ethyl, isopropyl and t-butyl.

In some embodiments, Y¹ as alkylamino can form acid addition salts.Suitable acids are well known in the art, for example hydrochloric acidand acetic acid.

In some embodiments, X is selected from cyano, 5-tetrazolyl,alkylsulfonylcarbamyl having a C₁₋₆ alkyl moiety thereof and a group—CO, Y, wherein Y is —OR¹ and R¹ is hydrogen, C₁₋₆ alkyl or benzyl, or Yis —NR²R³ where R² and R³ are independently hydrogen or alkyl of 1 to 4carbon atoms.

In some embodiments, X is carboxyl.

In some embodiments, the compound of Formula (I) is tucaresol.

Tucaresol, 4-[(2-formyl-3-hydroxy-phenoxy)methyl]benzoic acid, and itsanalog are immune modulators. These compounds can he readily preparedaccording to methods and procedures details in U.S. Pat. No. 4,535,183,which is incorporated herein by reference in its entirety. In someembodiments, these compounds can enhance co-stimulatory signaling toCD4-positive and CD8-positive T-cells, leading to tumor-cell-killing. Insome embodiments, these compounds can provide a costimulatory signal toCD4⁺ T-cells and CD8⁺ T-cells, activating Na⁺ and K⁺ transport,converging with T-cell receptor (TCR) signaling at the level of the MAPkinase ERK-2, and priming for increased intensity of calcium signaling.In some embodiments, these compounds can be biologically active as animmunopotentiator, favoring a Th1 response in patients with malignantmelanoma.

Plinabulin,(3Z,6Z)-3-Benzylidene-6-{[5-(2-methyl-2-propanyl)-1H-imidazol-4-yl]methylene}-2,5-piperazinedione,is a synthetic analog of the natural compound phenylahistin. Plinabulincan be readily prepared according to methods and procedures detailed inU.S. Pat. Nos. 7,064,201 and 7,919,497, which are incorporated herein byreference in their entireties. In some embodiments, Plinabulin canefficiently promote antigen uptake and migration of dendritic cells tolymph nodes where tumor-specific antigens are presented by dendriticcells to primeimmune effector cells. Exposure of dendritic cells toPlinabulin can induce maturation of dendritic cells and significantlyincrease their capacity to prime T cells. In some embodiments,Plinabulin can mediate tumor size reduction through immune modulation ofthe tumor microenvironment to promote anti-tumor immune enhancingeffects. In some embodiments, substantial therapeutic synergies can beachieved when combining Plinabulin with immune checkpoint inhibitors.

100261 Some embodiments relate to the use of a compound of formula (I)in combination with one or more immune checkpoint inhibitors, such asinhibitors of CTLA4 (cytotoxic T lymphocyte antigen-4), PD1 (programmedcell death protein 1), PD-L1 (programmed cell death ligand 1),PD-L2(programmed cell death ligand 2), PD-L3(programmed cell deathligand 3), PD-L4(programmed cell death ligand 4), LAG-3(lymphocyteactivation gene-3), and TIM-3 (T cell immunoglobulin and mucinprotein-3). In some embodiments, the compound of Formula (I) istucaresol. In some embodiments, the immune checkpoint inhibitor is abinding ligand of PD-1. In some embodiments, the immune checkpointinhibitor is a binding ligand of CTLA-4. Some embodiments relate to theuse of a compound of Formula (I) in combination with plinabulin. Someembodiments relate to the use of a compound of Formula (I) incombination with one or more immune checkpoint inhibitor and plinabulin.Some embodiments relate to the use of tucaresol in combination withplinabulin. Some embodiments relate to the use of tucaresol incombination with one or more immune checkpoint inhibitor and plinabulin.Some embodiments relate to the use of a compound of Formula (I) incombination with a PD-1 inhibitor or PD-L1 inhibitor. Some embodimentsrelate to the use of a compound of Formula (I) in combination with aninhibitor of PD-1 or PD-L1 and an inhibitor of CTLA-4. Some embodimentsrelate to the use of a compound of Formula (I) in combination with aninhibitor of PD-1, an inhibitor of PD-L1, and an inhibitor of CTLA-4.

Some embodiments relate to the use of tucaresol in combination with aPD-1 inhibitor or PD-L1 inhibitor. In some embodiments, no otheradditional checkpoint inhibitors are administered. Having no otheradditional check point inhibitors in the treatment may help to achievean effective treatment with reduced or minimal toxicity. In someembodiments, no inhibitor of CTLA-4 is administered.

Some embodiments relate to the use of tucaresol in combination with aninhibitor of PD-1 or PD-L1, and an inhibitor of CTLA-4. In someembodiments, the inhibitor of CTLA-4 is administered at a dose of lessthan 3 mg/kg. When the dose of the CTLA-4 inhibitor is lower than theconventional dose used for treating cancer or tumor growth (e.g., 3mg/kg), the treatment using a combination of tucaresol, an inhibitor ofPD-1 or PD-L1, and an inhibitor of CTLA-4 can lead to an increasedefficacy with reduced toxicity.

Some embodiments relate to the use of tucaresol in combination with aninhibitor of PD-1 or PD-L1 and an inhibitor of CTLA-4, wherein theinhibitor of CTLA-4 is administered at a dose of about 3 mg/kg orgreater. When the dose of the CTLA-4 inhibitor is the conventional doseused for treating cancer or tumor growth (e.g., 3 mg/kg), the treatmentof using this combination can lead to an increased efficacy withoutincreasing toxicity.

PD-1 is a key immune checkpoint receptor expressed by activated T and Bcells and mediates immunosuppression. PD-1 is a member of the CD28family of receptors, which includes CD28, CTLA-4, ICOS, PD-1, and BTLA.The term “PD-1” as used herein includes human PD-1 (hPD-1), variants,isoforms, and species homologs of hPD-1, and analogs having at least onecommon epitope with hPD-1.

Various cell surface glycoprotein ligands for PD-1 have been identified,including PD-L1, PD-L2, PD-L3, and PD-L4, that are expressed onantigen-presenting cells as well as many human cancers and have beenshown to downregulate cell activation and cytokine secretion uponbinding to PD-1. The term “PD-L1” as used herein includes human PD-L1(hPD-L1), variants, isoforms, and species homologs of hPD-L1, andanalogs having at least one common epitope with hPD-L1. The term “PD-L2”as used herein includes human PD-L2 (hPD-L2), variants, isoforms, andspecies homologs of hPD-L2, and analogs having at least one commonepitope with hPD-L2. The term “PD-L3” as used herein includes humanPD-L3 (hPD-L3), variants, isoforms, and species homologs of hPD-L3, andanalogs having at least one common epitope with hPD-L3. The term “PD-L4”as used herein includes human PD-L4 (hPD-L4), variants, isoforms, andspecies homologs of hPD-L4, and analogs having at least one commonepitope with hPD-L4.

CTLA-4 (cytotoxic T-lymphocyte-associated protein 4) is a proteinreceptor that, functioning as an immune checkpoint, downregulates theimmune system. CTLA4 is found on the surface of T cells, is also amember of the immunoglobulin (Ig) superfamily; CTLA-4 comprises a singleextracellular Ig domain. CTLA-4 transcripts have been found in T cellpopulations having cytotoxic activity, suggesting that CTLA-4 mightfunction in the cytolytic response.

The compound of Formula (I) described herein (e.g., tucaresol) can havea synergistic effect with immune checkpoint inhibitors such asPD-1/PD-L1 antibodies when used for activating the innate immune systemsuch as natural killer cells, mast cells, eosinophils, basophils; andthe phagocytic cells include macrophages, neutrophils, and dendriticcells. Activation of the innate immune system can be effective intreating cancer or inhibiting tumor growth.

The compound of Formula (I) described herein (e.g., tucaresol) can havea synergistic effect with immune checkpoint inhibitors such asPD-1/PD-L1 antibodies when used for inhibiting tumor growth. Inaddition, the compound of Formula (I) can also have a synergistic effectwith both the immune checkpoint inhibitors PD-1 antibody and CTLA-4antibody when used for inhibiting tumor growth. The compound of Formula(I) (e.g., tucaresol) generally can have superior anti-tumor propertiesover CTLA-4 antibody when used together with one or more other immunecheck point inhibitors and it has a better toxicity and safety profilethan CTLA-4 antibody. Therefore, tucaresol can be used as a superiorreplacement or supplement of CTLA-4 antibody in the chemotherapy.

Definitions

Unless defined otherwise, all technical and scientific terms used hereinhave the same meaning as is commonly understood by one of ordinary skillin the art to which this disclosure belongs, All patents, applications,published applications, and other publications are incorporated byreference in their entirety. In the event that there is a plurality ofdefinitions for a term herein, those in this section prevail unlessstated otherwise.

The term “pharmaceutically acceptable carrier” or “pharmaceuticallyacceptable excipient” includes any and all solvents, dispersion media,coatings, antibacterial and antifungal agents, isotonic and absorptiondelaying agents and the like. The use of such media and agents forpharmaceutically active substances is well known in the art. Exceptinsofar as any conventional media or agent is incompatible with theactive ingredient, its use in the therapeutic compositions iscontemplated. In addition, various adjuvants such as are commonly usedin the art may be included. Considerations for the inclusion of variouscomponents in pharmaceutical compositions are described, e.g., in Gilmanet al. (Eds.) (1990); Goodman and Gilman's: The Pharmacological Basis ofTherapeutics, 8th Ed., Pergamon Press, which is incorporated herein byreference in its entirety. The pharmaceutically acceptable excipient canbe a monosaccharide or monosaccharide derivative.

“Subject” as used herein, means a human or a non-human mammal, e.g., adog, a cat, a mouse, a rat, a cow, a sheep, a pig, a goat, a non-humanprimate or a bird, e.g., a chicken, as well as any other vertebrate orinvertebrate.

The term “mammal” is used in its usual biological sense. Thus, itspecifically includes, but is not limited to, primates, includingsimians (chimpanzees, apes, monkeys) and humans, cattle, horses, sheep,goats, swine, rabbits, dogs, cats, rodents, rats, mice, guinea pigs, orthe like.

An “effective amount” or a “therapeutically effective amount” as usedherein refers to an amount of a therapeutic agent that is effective torelieve, to some extent, or to reduce the likelihood of onset of, one ormore of the symptoms of a disease or condition, and can include curing adisease or condition.

“Treat,” “treatment,” or “treating,” as used herein refers toadministering a compound or pharmaceutical composition to a subject forprophylactic and/or therapeutic purposes. The term “prophylactictreatment” refers to treating a subject who does not yet exhibitsymptoms of a disease or condition, but who is susceptible to, orotherwise at risk of, a particular disease or condition, whereby thetreatment reduces the likelihood that the patient will develop thedisease or condition. The term “therapeutic treatment” refers toadministering treatment to a subject already suffering from a disease orcondition.

As used herein, the term “chemotherapeutic agent” refers to an agentthat reduces, prevents, mitigates, limits, and/or delays the growth ofmetastases or neoplasms, or kills neoplastic cells directly by necrosisor apoptosis of neoplasms or any other mechanism, or that can beotherwise used, in a pharmaceutically-effective amount, to reduce,prevent, mitigate, limit, and/or delay the growth of metastases orneoplasms in a subject with neoplastic disease. Chemotherapeutic agentsinclude but are not limited to, for example, fluoropyrimidines;pyrimidine nucleosides; purine nucleosides; anti-folates, platinum-basedagents; anthracyclines/anthracenediones; epipodophyllotoxins;camptothecins; hormones; hormonal complexes; antihormonals; enzymes,proteins, peptides and polyclonal and/or monoclonal antibodies; vincaalkaloids; taxanes; epothilones; antimicrotubule agents; alkylatingagents; antimetabolites; topoisomerase inhibitors; antivirals; andvarious other cytotoxic and cytostatic agents.

The term “pharmaceutically acceptable salt” refers to salts that retainthe biological effectiveness and properties of a compound and, which arenot biologically or otherwise undesirable for use in a pharmaceutical.In many cases, the compounds disclosed herein are capable of formingacid and/or base salts by virtue of the presence of amino and/orcarboxyl groups or groups similar thereto. Pharmaceutically acceptableacid addition salts can be formed with inorganic acids and organicacids. Inorganic acids from which salts can be derived include, forexample, hydrochloric acid, hydrobromic acid, sulfuric acid, nitricacid, phosphoric acid, and the like. Organic acids from which salts canbe derived include, for example, acetic acid, propionic acid, glycolicacid, pyruvic acid, oxalic acid, maleic acid, malonic acid, succinicacid, fumaric acid, tartaric acid, citric acid, benzoic acid, cinnamicacid, mandelic acid, methanesulfonic acid, ethanesulfonic acid,p-toluenesulfonic acid, salicylic acid, and the like. Pharmaceuticallyacceptable base addition salts can be formed with inorganic and organicbases. Inorganic bases from which salts can be derived include, forexample, sodium, potassium, lithium, ammonium, calcium, magnesium, iron,zinc, copper, manganese, aluminum, and the like; particularly preferredare the ammonium, potassium, sodium, calcium and magnesium salts.Organic bases from which salts can be derived include, for example,primary, secondary, and tertiary amines, substituted amines includingnaturally occurring substituted amines, cyclic amines, basic ionexchange resins, and the like, specifically such as isopropylamine,methylamine, diethylamine, triethylamine, tripropylamine, andethanolamine. Many such salts are known in the art, as described in WO87/05297, Johnston et al., published Sep. 11, 1987 (incorporated byreference herein in its entirety).

As used herein, “C_(a) to C_(b)” or “C_(a-b),” in which “a” and “b” areintegers refer to the number of carbon atoms in the specified group.That is, the group can contain from “a” to “b”, inclusive, carbon atoms.Thus, for example, a “C₁ to C₄ alkyl” or “C₁₋₄ alkyl” group refers toall alkyl groups having from 1 to 4 carbons, that is, CH₃—, CH₃CH₂—,CH₃CH₂CH₂—, (CH₃)₂CH—, CH₃CH₂CH₂CH₂—, CH₃CH₂CH(CH₃)— and (CH₃)₃C—.

The term “halogen” or “halo,” as used herein, means any one of theradio-stable atoms of column 7 of the Periodic Table of the Elements,e.g., fluorine, chlorine, bromine, or iodine, with fluorine and chlorinebeing preferred.

As used herein, “alkyl” refers to a straight or branched hydrocarbonchain that is fully saturated (i.e., contains no double or triplebonds). The alkyl group may have 1 to 20 carbon atoms (whenever itappears herein, a numerical range such as “1 to 20” refers to eachinteger in the given range; e.g., “l to 20 carbon atoms” means that thealkyl group may consist of 1 carbon atom, 2. carbon atoms, 3 carbonatoms, etc., up to and including 20 carbon atoms, although the presentdefinition also covers the occurrence of the term “alkyl” where nonumerical range is designated). The alkyl group may also be a mediumsize alkyl having 1 to 9 carbon atoms. The alkyl group could also be alower alkyl having 1 to 4 carbon atoms. The alkyl group may bedesignated as “C₁₋₄ alkyl” or similar designations. By way of exampleonly, “C₁₋₄ alkyl” indicates that there are one to four carbon atoms inthe alkyl chain, i.e., the alkyl chain is selected from the groupconsisting of methyl, ethyl, propyl, iso-propyl, n-butyl, i so-butyl,sec-butyl, and t-butyl. Typical alkyl groups include, but are in no waylimited to, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tertiarybutyl, pentyl, hexyl, and the like.

As used herein, “alkoxy” refers to the formula —OR wherein R is an alkylas is defined above, such as “C₁₋₉ alkoxy”, including but not limited tomethoxy, ethoxy, n-propoxy, 1-methylethoxy (isopropoxy), n-butoxy,iso-butoxy, sec-butoxy, and tert-butoxy, and the like.

As used herein, “alkylthio” refers to the formula —SR wherein R is analkyl as is defined above, such as “C₁₋₉ alkylthio” and the like,including but not limited to methylmercapto, ethylmercapto,n-propylmercapto, 1-methylethyltnercapto (isopropylmercapto),n-butylmercapto, iso-butylmercapto, sec-butylmercapto,tert-butylmercapto, and the like,

As used herein, “aryl” refers to an aromatic ring or ring system (i.e.,two or more fused rings that share two adjacent carbon atoms) containingonly carbon in the ring backbone. When the aryl is a ring system, everyring in the system is aromatic. The aryl group may have 6 to 18 carbonatoms, although the present definition also covers the occurrence of theterm “aryl” where no numerical range is designated. In some embodiments,the aryl group has 6 to 10 carbon atoms. The aryl group may bedesignated as “C₆₋₁₀ aryl,” “C₆ or C₁₀ aryl,” or similar designations.Examples of aryl groups include, but are not limited to, phenyl,naphthyl, azulenyl, and anthracenyl.

As used herein, “heteroaryl” refers to an aromatic ring or ring systemtwo or more fused rings that share two adjacent atoms) that contain(s)one or more heteroatoms, that is, an element other than carbon,including but not limited to, nitrogen, oxygen and sulfur, in the ringbackbone. When the heteroaryl is a ring system, every ring in the systemis aromatic. The heteroaryl group may have 5-18 ring members (i.e., thenumber of atoms making up the ring backbone, including carbon atoms andheteroatoms), although the present definition also covers the occurrenceof the term “heteroaryl” where no numerical range is designated. In someembodiments, the heteroaryl group has 5 to 10 ring members or 5 to 7ring members. The heteroaryl group may be designated as “5-7 memberedheteroaryl,” “5-10 membered heteroaryl,” or similar designations.Examples of heteroaryl rings include, but are not limited to, furyl,thienyl, phthalazinyl, pyrrolyl, oxazolyl, thiazolyl, imidazolyl,pyrazolyl, isoxazolyl, isothiazolyl, triazolyl, thiadiazolyl, pyridinyl,pyridazinyl, pyrimidinyl, pyrazinyl, triazinyl, quinolinyl,isoquinlinyl, benzimidazolyl, benzoxazolyl, benzothiazolyl, indolyl,isoindolyl, and benzothienyl.

As used herein, “acyl” refers to —C(═O)R, wherein R is hydrogen, C₁₋₆alkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl, C₃₋₇ carbocyclyl, C₆₋₁₀ aryl, 5-10membered heteroaryl, and 5-10 membered heterocyclyl, as defined herein.Non-limiting examples include formyl, acetyl, propanoyl, benzoyl, andacryl.

An “O-carboxy” group refers to a “—OC(═O)R” group in which R is selectedfrom hydrogen, C₁₋₆ alkyl, C₂₋₆ alkenyl; C₂₋₆ alkynyl, C₃₋₇ carbocyclyl,C₆₋₁₀ aryl, 5-10 membered heteroaryl, and 5-10 membered heterocyclyl, asdefined herein.

A “C-carboxy” group refers to a “—C(═O)OR” group in which R is selectedfrom hydrogen, C₁₋₆ alkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl, C₃₋₇ carbocyclyl,C₆₋₁₀ aryl, 5-10 membered heteroatyl, and 5-10 membered heterocyclyl, asdefined herein. A non-limiting example includes carboxyl (i.e.,—C(═O)OH).

A “sulfonyl” group refers to an “—SO₂R” group in which R is selectedfrom hydrogen, C₁₋₆ alkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl, C₃₋₇ carbocyclyl,C₆₋₁₀ aryl, 5-10 membered heteroaryl, and 5-10 membered heterocyclyl, asdefined herein.

An “amino” group refers to a “—NR_(A)R_(B)” group in which R_(A) andR_(B) are each independently selected from hydrogen, C₁₋₆ alkyl, C₂₋₆alkenyl, C₂₋₆ alkynyl, C₃₋₇ carbocyclyl, C₆₋₁₀ aryl, 5-10 memberedheteroaryl, and 5-10 membered heterocyclyl, as defined herein. Anon-limiting example includes free amino (i.e., —NH₂).

An “aminoalkyl” group refers to an amino group connected via an alkylenegroup.

As used herein, a substituted group is derived from the unsubstitutedparent group in which there has been an exchange of one or more hydrogenatoms for another atom or group. Unless otherwise indicated, when agroup is deemed to be “substituted,” it is meant that the group issubstituted with one or more subsitutents independently selected fromC₁-C₆ alkyl, C₁-C₆ alkenyl, C₁-C₆ alkynyl, C₁-C₆ heteroalkyl, C₃-C₇carbocyclyl (optionally substituted with halo, C₁-C₆ alkyl, C₁-C₆alkoxy, C₁-C₆ haloalkyl, and C₁-C₆ haloalkoxy),C₃-C₇-carbocyclyl-C₁-C₆-alkyl (optionally substituted with halo, C₁-C₆alkyl, C₁-C₆ alkoxy, C₁-C₆ haloalkyl, and C₁-C₆ haloalkoxy), 5-10membered heterocyclyl (optionally substituted with halo, C₁-C₆ alkyl,C₁-C₆ alkoxy, C₁-C₆ haloalkyl, and C₁-C₆ haloalkoxy), 5-10 memberedheterocyclyl-C₁-C₆-alkyl(optionally substituted with halo, C₁-C₆ alkyl,C₁-C₆ alkoxy, C₁-C₆ haloalkyl, and C₁-C₆ haloalkoxy), aryl (optionallysubstituted with halo, C₁-C₆ alkyl, C₁-C₆ alkoxy, C₁-C₆ haloalkyl, andC₁-C₆ haloalkoxy), aryl(C₁-C₆)alkyl (optionally substituted with halo,C₁-C₆ alkyl, C₁-C₆ alkoxy, C₁-C₆ haloalkyl, and C₁-C₆ haloalkoxy), 5-10membered heteroaryl (optionally substituted with halo, C₁-C₆ alkyl,C₁-C₆ alkoxy, C₁-C₆ haloalkyl, and C₁-C₆ haloalkoxy), 5-10 memberedheteroaryl(C₁-C₆)alkyl (optionally substituted with halo, C₁-C₆ alkyl,C₁-C₆ alkoxy, C₁-C₆ haloalkyl, and C₁-C₆ halloalkoxy), halo, cyano,hydroxy, C₁-C₆ alkoxy, C₁-C₆ alkoxy(C₁-C₆)alkyl (i.e., ether), aryloxy,sulfhydryl (mercapto), halo(C₁-C₆)alkyl (e.g., —CF₃), halo(C₁-C₆)alkoxy(e.g., C₁-C₆ alkylthio, arylthio, amino, amino(C₁-C₆)alkyl, nitro,O-carbamyl, N-carbamyl, O-thiocathamyl, N-thiocarbamyl, C-amido,N-amido, S-sulfonamido, N-sulfonamido, C-carboxy, O-carboxy, acyl,cyanato, isocyanate, thiocyanate, isothiocyanate, sulfinyl, sulfonyl,and oxo (═O). Wherever a group is described as “optionally substituted”that group can be substituted with the above substituents.

It is to be understood that certain radical naming conventions caninclude either a mono-radical or a di-radical, depending on the context.For example, where a substituent requires two points of attachment tothe rest of the molecule, it is understood that the substituent is adi-radical. For example, a substituent identified as alkyl that requirestwo points of attachment includes di-radicals such as —CH₂—, —CH₂CH₂—,—CH₂CH(CH₃)CH₂—, and the like. Other radical naming conventions clearlyindicate that the radical is a di-radical such as “alkylene” or“alkenylene.”

Wherever a substituent is depicted as a di-radical (i.e., has two pointsof attachment to the rest of the molecule), it is to be understood thatthe substituent can be attached in any directional configuration unlessotherwise indicated. Thus, for example, a substituent depicted as -AE-or

includes the substituent being oriented such that the A is attached atthe leftmost attachment point of the molecule as well as the case inwhich A is attached at the rightmost attachment point of the molecule.

Unless defined otherwise, all technical and scientific terms used hereinhave the same meaning as commonly understood by one of ordinary skill inthe art to which the embodiments belong. Although any methods andmaterials similar or equivalent to those described herein can also beused in the practice or testing of the embodiments, the preferredmethods and materials are now described. All publications mentionedherein are incorporated herein by reference to disclose and describe themethods and/or materials in connection with which the publications arecited.

Administration and Pharmaceutical Compositions

Some embodiments relate to a pharmaceutical composition including acompound of Formula (I) described herein and one or more immunecheckpoint inhibitor. In some embodiments, the compound of Formula (I)is tucaresol. In some embodiments, the composition described hereinfurther includes plinabulin. In some embodiments, the immune checkpointinhibitor is an inhibitor of PD-1, PD-L1, PD-L2, PD-L3, PD-L4, CTLA-4,LAG3, B7-H3, B7-H4, KIR or TIM3. In some embodiments, the immunecheckpoint inhibitor is a PD-1 inhibitor. In some embodiments, theimmune checkpoint inhibitor is a binding ligand of PD-L1. In someembodiments, the immune checkpoint inhibitor is a PD-L1 inhibitor. Insome embodiments, the immune checkpoint inhibitor is a PD-L2 inhibitoror a combined PD-L1/PD-L2 inhibitor. In some embodiments, the immunecheckpoint inhibitor is a CTLA-4 inhibitor.

In some embodiments, the composition described herein includes acompound of Formula (I), a first immune checkpoint inhibitor and asecond immune checkpoint inhibitor, wherein the first immune checkpointinhibitor is different from the second immune checkpoint inhibitor. Insome embodiments, the first and the second immune checkpoint inhibitoris independently selected from an inhibitor of PD-1, PD-L1, PD-L2,PD-L3, PD-L4, CTLA-4, LAG-3, B7-H3, B7-H4, KIR or TIM3. In someembodiments, the first immune checkpoint inhibitor is a PD-1 inhibitor,and the second immune checkpoint inhibitor is a CTLA-4 inhibitor. Insome embodiments, the first immune checkpoint inhibitor is a PD-L1inhibitor, and the second immune checkpoint inhibitor is a CTLA-4inhibitor. In some embodiments, the first immune checkpoint inhibitor isa PD-L2 inhibitor, and the second immune checkpoint inhibitor is aCTLA-4 inhibitor.

In some embodiments, the composition described herein can include thecompound of formula (I) and an inhibitor of PD1. In some embodiments,the composition described herein can include the compound of formula (I)and an inhibitor of PD-L1. In some embodiments, the compositiondescribed herein can include the compound of formula (I), an inhibitorof PD-1 or PD-L1, and an inhibitor of CTLA-4. In some embodiments, thePD-1 inhibitor is pembrolizumab. In some embodiments, the PD-1 inhibitoris nivolumab. In some embodiments, the PD-L1 inhibitor is atezolizumab.In some embodiments, the CTLA-4 inhibitor is ipilimumab.

In some embodiments, the immune checkpoint inhibitor can be a smallpeptide agent that can inhibit T cell regulation function. In someembodiments, the immune checkpoint inhibitor can be a small molecule(e.g. less than 500 Daltons) that can inhibit T cell regulationfunction. In some embodiments, the immune checkpoint inhibitor can be amolecule providing co-stimulation of T-cell activation. In someembodiments, the immune checkpoint inhibitor can be a molecule providingco-stimulation of natural killer cell activation. In some embodiments,the immune checkpoint inhibitor can be an antibody. In some embodiments,the immune checkpoint inhibitor is a PD-1 antibody. In some embodiments,the immune checkpoint inhibitor is a PD-L1 antibody. In someembodiments, the immune checkpoint inhibitor is a PD-L2 antibody. Insome embodiments, the immune checkpoint inhibitor is a PD-L3 antibody.In some embodiments, the immune checkpoint inhibitor is a PD-L4antibody. In some embodiments, the immune checkpoint inhibitor is aCTLA-4 antibody. In some embodiments, the immune checkpoint inhibitor isan antibody of CTLA-4, LAG3, B7-H3, B7-H4, KIR, or TIM3.

The antibody can be selected from α-CD3-APC, α-CD3-APC-H7, α-CD4-ECD,α-CD4-PB, α-CD8-PE-Cy7, α-CD-8-PerCP-Cy5.5, α-CD11c-APC, α-CD11b-PE-Cy7,α-CD11b-AF700, α-CD14-FITC, α-CD16-PB, α-CD19-AF780, α-CD19-AF700,α-CD20-PO, α-CD25-PE-Cy7, α-CD40-APC, α-CD45-Biotin, Streptavidin-BV605,α-CD62L-ECD, α-CD69-APC-Cy7, α-CD80-FITC, α-CD83-Biotin,Streptavidin-PE-Cy7, α-CD86-PE-Cy7, α-CD86-PE, α-CD123-PE, α-CD154-PE,α-CD161-PE, α-CTLA4-PE-Cy7, α-FoxP3-AF488 (clone 259D),IgG1-isotype-AF488, α-ICOS (CD278)-PE, α-HLA-A2-PE, α-HLA-DR-PB,α-HLA-DR-PerCPCy5.5, α-PD1-APC, VISTA, co-stimulatory molecule OX40, andCD137.

A variety of antibodies (Abs) can be used in the composition describedherein, including antibodies having high-affinity binding to PD-1,PD-L1, PD-L2, PD-L3, or PD-L4. Human mAbs (HuMAbs) that bindspecifically to PD-1 (e.g., bind to human PD-1 and may cross-react withPD-1 from other species, such as cynomolgus monkey) with high affinityhave been disclosed in U.S. Pat. No, 8,008,449, which is incorporatedherein by reference in its entirety. HuMAbs that bind specifically toPD-L1 with high affinity have been disclosed in U.S. Pat. No. 7,943,743,which is incorporated herein by reference in its entirety. Otheranti-PD-1 mAbs have been described in, for example, U.S. Pat. Nos.6,808,710, 7,488,802 and 8,168,757, and PCT Publication No. WO2012/145493, all of which are incorporated herein by reference in theirentireties, Anti-PD-L1 mAbs have been described in, for example, U.S.Pat. Nos. 7,635,757 and 8,217,149, U.S. Publication No. 2009/0317368,and PCT Publication Nos. WO 2011/066389 and WO 2012/14549, all of whichare incorporated herein by reference in their entireties.

In some embodiments, the anti-PD-1 HuMAbs can be selected from 17D8,2D3, 4H1, 5C4 (also referred to herein as nivolumab), 4A1 1, 7D3 and5F4, all of which are described in U.S. Pat. No. 8,008,449. In someembodiments, the anti-PD-1 HuMAbs can be selected from 3G10, 12A4 (alsoreferred to herein as BMS-936559), 10A5, 5F8, 10H10, 1B12, 7H1, 1 1E6,12B7, and 13G4, all of which are described in U.S. Pat. No. 7,943,743.

Some embodiments relate to a pharmaceutical composition comprising acompound of Formula (I) and plinabulin. In some embodiments, thecompound of Formula (I) is tucaresol.

In some embodiments, the composition can further include one or morepharmaceutically acceptable diluents. In some embodiments, thepharmaceutically acceptable diluent can include Kolliphor HS15®(Polyoxyl (15)-hydroxystearate). In some embodiments, thepharmaceutically acceptable diluent can include propylene glycol. Insome embodiments, the pharmaceutically acceptable diluents can includekolliphor and propylene glycol. In some embodiments, thepharmaceutically acceptable diluents can include kolliphor and propyleneglycol, wherein the kolliphor is about 40% by weight and propyleneglycol is about 60% by weight based on the total weight of the diluents.In some embodiments, the composition can further include one or moreother pharmaceutically acceptable excipients.

Standard pharmaceutical formulation techniques can be used to make thepharmaceutical compositions described herein, such as those disclosed inRemington's The Science and Practice of Pharmacy, 21st Ed., LippincottWilliams & Wilkins (2005), incorporated herein by reference in itsentirety. Accordingly, some embodiments include pharmaceuticalcompositions comprising: (a) a safe and therapeutically effective amountof Plinabulin or pharmaceutically acceptable salts thereof; (b) animmune checkpoint inhibitor and (c) a pharmaceutically acceptablecarrier, diluent, excipient or combination thereof.

Other embodiments include co-administering a compound of Formula (I) andone or more immune checkpoint inhibitor in separate compositions. Insome embodiments, the compound of Formula (I) is tucaresol. Thus, someembodiments include a first pharmaceutical compositions comprising: (a)a safe and therapeutically effective amount of the compound of Formula(I) or pharmaceutically acceptable salts thereof and (b) apharmaceutically acceptable carrier, diluent, excipient or combinationthereof; and a second pharmaceutical composition comprising: (a) one ormore immune checkpoint inhibitor and (b) a pharmaceutically acceptablecarrier, diluent, excipient or combination thereof.

Other embodiments include co-administering a compound of Formula (I) andplinabulin in separate compositions. In some embodiments, the compoundof Formula (I) is tucaresol. Thus, some embodiments include a firstpharmaceutical compositions comprising: (a) a safe and therapeuticallyeffective amount of the compound of Formula (I) or pharmaceuticallyacceptable salts thereof and (b) a pharmaceutically acceptable carrier,diluent, excipient or combination thereof; and a second pharmaceuticalcomposition comprising: (a) plinabulin and (b) a pharmaceuticallyacceptable carrier, diluent, excipient or combination thereof.

Other embodiments include co-administering a compound of Formula (I),one or more immune checkpoint inhibitor, and plinabulin in separatecompositions. In some embodiments, the compound of Formula (I) istucaresol. Thus, some embodiments include a first pharmaceuticalcompositions comprising: (a) a safe and therapeutically effective amountof the compound of Formula (I) or pharmaceutically acceptable saltsthereof and (b) a pharmaceutically acceptable carrier, diluent,excipient or combination thereof; and a second pharmaceuticalcomposition comprising: (a) one or more immune checkpoint inhibitor and(b) a pharmaceutically acceptable carrier, diluent, excipient orcombination thereof; and a third pharmaceutical composition comprising:(a) plinabulin and (b) a pharmaceutically acceptable carrier, diluent,excipient or combination thereof.

Administration of the pharmaceutical compositions described herein canbe via any of the accepted modes of administration for agents that servesimilar utilities including, but not limited to, orally, sublingually,buccally, subcutaneously, intravenously, intranasally, topically,transdermally, intradermally, intraperitoneally, intramuscularly,intrapulmonarilly, vaginally, rectally, or intraocularly. Oral andparenteral administrations are customary in treating the indicationsthat are the subject of the preferred embodiments.

The term “pharmaceutically acceptable carrier” or “pharmaceuticallyacceptable excipient” includes any and all solvents, dispersion media,coatings, antibacterial and antifungal agents, isotonic and absorptiondelaying agents and the like. The use of such media and agents forpharmaceutically active substances is well known in the art. Exceptinsofar as any conventional media or agent is incompatible with theactive ingredient, its use in the therapeutic compositions iscontemplated. In addition, various adjuvants such as are commonly usedin the art may be included. Considerations for the inclusion of variouscomponents in pharmaceutical compositions are described, e.g., in Gilmanet al. (Eds.) (1990); Goodman and Gilman's: The Pharmacological Basis ofTherapeutics, 8th Ed., Pergamon Press, which is incorporated herein byreference in its entirety.

Some examples of substances, which can serve aspharmaceutically-acceptable carriers or components thereof, are sugars,such as lactose, glucose and sucrose; starches, such as corn starch andpotato starch; cellulose and its derivatives, such as sodiumcarboxymethyl cellulose, ethyl cellulose, and methyl cellulose; powderedtragacanth; malt; gelatin; talc; solid lubricants, such as stearic acidand magnesium stearate; calcium sulfate; vegetable oils, such as peanutoil, cottonseed oil, sesame oil, olive oil, corn oil and oil oftheobroma; polyols such as propylene glycol, glycerine, sorbitol,mannitol, and polyethylene glycol; alginic acid; emulsifiers, such asthe TWEENS; wetting agents, such sodium lauryl sulfate; coloring agents;flavoring agents; tableting agents, stabilizers; antioxidants;preservatives; pyrogen-free water; isotonic saline; and phosphate buffersolutions.

The compositions described herein are preferably provided in unit dosageform. As used herein, a “unit dosage form” is a composition containingan amount of a compound or composition that is suitable foradministration to an animal, preferably mammal subject, in a singledose, according to good medical practice. The preparation of a single orunit dosage form however, does not imply that the dosage form isadministered once per day or once per course of therapy. Such dosageforms are contemplated to be administered once, twice, thrice or moreper day and may be administered as infusion over a period of time (e.g.,from about 30 minutes to about 2-6 hours), or administered as acontinuous infusion, and may be given more than once during a course oftherapy, although a single administration is not specifically excluded.The skilled artisan will recognize that the formulation does notspecifically contemplate the entire course of therapy and such decisionsare left for those skilled in the art of treatment rather thanformulation.

The compositions useful as described above may be in any of a variety ofsuitable forms for a variety of routes for administration, for example,for oral, sublingual, buccal, nasal, rectal, topical (includingtransdermal and intradermal), ocular, intracerebral, intracranial,intrathecal, intra-arterial, intravenous, intramuscular, or otherparental routes of administration. The skilled artisan will appreciatethat oral and nasal compositions include compositions that areadministered by inhalation, and made using available methodologies.Depending upon the particular route of administration desired, a varietyof pharmaceutically-acceptable carriers well-known in the art may beused. Pharmaceutically-acceptable carriers include, for example, solidor liquid fillers, diluents, hydrotropies, surface-active agents, andencapsulating substances. Optional pharmaceutically-active materials maybe included, which do not substantially interfere with the inhibitoryactivity of the compound or composition. The amount of carrier employedin conjunction with the compound or composition is sufficient to providea practical quantity of material for administration per unit dose of thecompound. Techniques and compositions for making dosage forms useful inthe methods described herein are described in the following references,all incorporated by reference herein: Modern Pharmaceutics, 4th Ed.,Chapters 9 and 10 (Banker & Rhodes, editors, 2002); Lieberman et al.,Pharmaceutical Dosage Forms: Tablets (1989); and Ansel, Introduction toPharmaceutical Dosage Forms 8th Edition (2004).

Various oral dosage forms can be used, including such solid forms astablets, capsules (e.g. solid gel capsules and liquid gel capsules),granules and bulk powders. Tablets can be compressed, tablet triturates,enteric-coated, sugar-coated, film-coated, or multiple-compressed,containing suitable binders, lubricants, diluents, disintegratingagents, coloring agents, flavoring agents, flow-inducing agents, andmelting agents. Liquid oral dosage forms include aqueous solutions,emulsions, suspensions, solutions and/or suspensions reconstituted fromnon-effervescent granules, and effervescent preparations reconstitutedfrom effervescent granules, containing suitable solvents, preservatives,emulsifying agents, suspending agents, diluents, sweeteners, meltingagents, coloring agents and flavoring agents.

The pharmaceutically-acceptable carriers suitable for the preparation ofunit dosage forms for peroral administration is well-known in the art.Tablets typically comprise conventional pharmaceutically-compatibleadjuvants as inert diluents, such as calcium carbonate, sodiumcarbonate, mannitol, lactose and cellulose; binders such as starch,gelatin and sucrose; disintegrants such as starch, alginic acid andcroscarmelose; lubricants such as magnesium stearate, stearic acid andtalc. Glidants such as silicon dioxide can be used to improve flowcharacteristics of the powder mixture. Coloring agents, such as the FD&Cdyes, can be added for appearance. Sweeteners and flavoring agents, suchas aspartame, saccharin, menthol, peppermint, and fruit flavors, areuseful adjuvants for chewable tablets. Capsules typically comprise oneor more solid diluents disclosed above. The selection of carriercomponents depends on secondary considerations like taste, cost, andshelf stability, which are not critical, and can be readily made by aperson skilled in the art.

Peroral compositions also include liquid solutions, emulsions,suspensions, and the like. The pharmaceutically-acceptable carrierssuitable for preparation of such compositions are well known in the art.Typical components of carriers for syrups, elixirs, emulsions andsuspensions include ethanol, glycerol, propylene glycol, polyethyleneglycol, liquid sucrose, sorbitol and water. For a suspension, typicalsuspending agents include methyl cellulose, sodium carboxymethylcellulose, AVICEL RC-591, tragacanth and sodium alginate; typicalwetting agents include lecithin and polysorbate 80; and typicalpreservatives include methyl paraben and sodium benzoate. Peroral liquidcompositions may also contain one or more components such as sweeteners,flavoring agents and colorants disclosed above.

Such compositions may also be coated by conventional methods, typicallywith pH or time-dependent coatings, such that the subject composition isreleased in the gastrointestinal tract in the vicinity of the desiredtopical application, or at various times to extend the desired action.Such dosage forms typically include, but are not limited to, one or moreof cellulose acetate phthalate, polyvinylacetate phthalate,hydroxypropyl methyl cellulose phthalate, ethyl cellulose, Eudragitcoatings, waxes and shellac.

Compositions described herein may optionally include other drug actives.

Other compositions useful for attaining systemic delivery of the subjectcompounds include sublingual, buccal and nasal dosage forms. Suchcompositions typically comprise one or more of soluble filler substancessuch as sucrose, sorbitol and mannitol; and binders such as acacia,microcrystalline cellulose, carboxymethyl cellulose and hydroxypropylmethyl cellulose. Glidants, lubricants, sweeteners, colorants,antioxidants and flavoring agents disclosed above may also be included.

A liquid composition, which is formulated for topical ophthalmic use, isformulated such that it can be administered topically to the eye. Thecomfort may be maximized as much as possible, although sometimesformulation considerations (e.g. drug stability) may necessitate lessthan optimal comfort, In the case that comfort cannot be maximized, theliquid may be formulated such that the liquid is tolerable to thepatient for topical ophthalmic use. Additionally, an ophthalmicallyacceptable liquid may either be packaged for single use, or contain apreservative to prevent contamination over multiple uses.

For ophthalmic application, solutions or medicaments are often preparedusing a physiological saline solution as a major vehicle. Ophthalmicsolutions may preferably be maintained at a comfortable pH with anappropriate buffer system. The formulations may also containconventional, pharmaceutically acceptable preservatives, stabilizers andsurfactants.

Preservatives that may be used in the pharmaceutical compositionsdisclosed herein include, but are not limited to, benzalkonium chloride,PHMB, chlorobutanol, thimerosal, phenylmercuric, acetate andphenylmercuric nitrate. A useful surfactant is, for example, Tween 80.Likewise, various useful vehicles may be used in the ophthalmicpreparations disclosed herein. These vehicles include, but are notlimited to, polyvinyl alcohol, povidone, hydroxypropyl methyl cellulose,poloxamers, carboxymethyl cellulose, hydroxyethyl cellulose and purifiedwater.

Tonicity adjustors may be added as needed or convenient. They include,but are not limited to, salts, particularly sodium chloride, potassiumchloride, mannitol and glycerin, or any other suitable ophthalmicallyacceptable tonicity adjustor.

Various buffers and means for adjusting pH may be used so long as theresulting preparation is ophthalmically acceptable. For manycompositions, the pH will be between 4 and 9. Accordingly, buffersinclude acetate buffers, citrate buffers, phosphate buffers and boratebuffers. Acids or bases may be used to adjust the pH of theseformulations as needed.

Ophthalmically acceptable antioxidants include, but are not limited to,sodium metabisulfite, sodium thiosulfate, acetylcysteine, butylatedhydroxyanisole and butylated hydroxytoluene.

Other excipient components, which may be included in the ophthalmicpreparations, are chelating agents. A useful chelating agent is edetatedisodium, although other chelating agents may also be used in place orin conjunction with it.

For topical use, creams, ointments, gels, solutions or suspensions,etc., containing the composition disclosed herein are employed. Topicalformulations may generally be comprised of a pharmaceutical carrier,co-solvent, emulsifier, penetration enhancer, preservative system, andemollient.

For intravenous administration, the compositions described herein may bedissolved or dispersed in a pharmaceutically acceptable diluent, such asa saline or dextrose solution. Suitable excipients may be included toachieve the desired pH, including but not limited to NaOH, sodiumcarbonate, sodium acetate, HCl, and citric acid. In various embodiments,the pH of the final composition ranges from 2 to 8, or preferably from 4to 7. Antioxidant excipients may include sodium bisulfite, acetonesodium bisulfite, sodium formaldehyde, sulfoxvlate, thiourea, and EDTA.Other non-limiting examples of suitable excipients found in the finalintravenous composition may include sodium or potassium phosphates,citric acid, tartaric acid, gelatin, and carbohydrates such as dextrose,mannitol, and dextran. Further acceptable excipients are described inPowell, et al., Compendium of Excipients for Parenteral Formulations,PDA J Pharm Sci and Tech 1998, 52 238-311 and Nema et al., Excipientsand Their Role in Approved Injectable Products: Current Usage and FutureDirections, PDA J Pharm Sci and Tech 2011, 65 287-332, both of which areincorporated herein by reference in their entirety. Antimicrobial agentsmay also be included to achieve a bacteriostatic or fungistaticsolution, including but not limited to phenylmercuric nitrate,thimerosal, benzethonium chloride, benzalkonium chloride, phenol,cresol, and chlorobutanol.

The compositions for intravenous administration may be provided tocaregivers in the form of one more solids that are reconstituted with asuitable diluent such as sterile water, saline or dextrose in watershortly prior to administration. In other embodiments, the compositionsare provided in solution ready to administer parenterally. In stillother embodiments, the compositions are provided in a solution that isfurther diluted prior to administration. In embodiments that includeadministering a combination of a compound described herein and anotheragent, the combination may be provided to caregivers as a mixture, orthe caregivers may mix the two agents prior to administration, or thetwo agents may be administered separately.

In some embodiments, the compositions described herein can be used incombination with other therapeutic agents. In some embodiments, thecompositions described herein can be administered or used in combinationwith treatments such as chemotherapy, radiation, and biologic therapies.

Method of Treatment

Some embodiments relate to a method for treating cancer using thepharmaceutical composition described herein to a subject in needthereof. Some embodiments relate to a method for treating cancer,comprising co-administering a compound of Formula (I) described hereinand one or more immune checkpoint inhibitor to a subject in needthereof. Some embodiments relate to a method for treating cancer,comprising co-administering a compound of Formula (I), one or moreimmune checkpoint inhibitor, and plinabulin to a subject in needthereof. Some embodiments relate to a method for treating cancer,comprising co-administering a compound of Formula (I) and plinabulin toa subject in need thereof. In some embodiments, the compound of Formula(I) is tucaresol. In some embodiments, the subject can be an animal,e.g., a mammal, a human, in some embodiments, the subject is a human.

Some embodiments relate to methods of providing co-stimulation of T-cellactivation against cancer by co-administering a compound of formula (I),one or more immune checkpoint inhibitor. Some embodiments relate tomethods of providing co-stimulation of natural killer cells againstcancer by co-administering a compound of formula (I), one or more immunecheckpoint inhibitor. In some embodiments, the compound of Formula (I)is tucaresol.

In some embodiments, the cancer comprises cancer cells expressing abinding ligand of PD-1. In some embodiments, the binding ligand of PD-1is PD-L1. In some embodiments, the binding ligand of PD-1 is PD-L2.

In some embodiments, the method of treating cancer described hereinfurther includes identifying cancer cells expressing a binding ligand ofPD-1. In some embodiments, the method of treating cancer describedherein further includes identifying cancer cells expressing PD-L1. Insome embodiments, the method of treating cancer described herein furtherincludes identifying cancer cells expressing PD-L2. In some embodiments,the method of treating cancer described herein further includesidentifying cancer cells expressing PD-L3 or PD-L4.

In some embodiments, identifying cancer cells expressing a bindingligand of PD-1 includes using an assay to detect the presence of thebinding ligand. Examples of applicable assay include but are not limitedto PD-L1 IHC 22C3 pharmDx kit and PD-L1 IHC 28-8 pharmDx available fromDako.

In some embodiments, the cancer comprises cancer calls expressing abinding ligand of CTLA-4. In some embodiments, the binding ligand ofCTLA-4 is B7.1 or B7.2.

In some embodiments, the method of treating cancer described hereinfurther includes identifying cancer cells expressing a binding ligand ofCTLA-4. In some embodiments, the method of treating cancer describedherein further includes identifying cancer cells expressing B7.1 orB7.2.

In some embodiments, the immune checkpoint inhibitor is nivolumab,pembrolizumab, pidilizumab, ipilimumab, dacarbazine, BMS 936559,a.tezolizumab, durvalitnuma.b, or any combinations thereof.

In some embodiments, cancer is head and neck cancer, lung cancer,stomach cancer, colon cancer, pancreatic cancer, prostate cancer, breastcancer, kidney cancer, bladder cancer, ovary cancer, cervical cancer,melanoma, glioblastoma, myeloma, lymphoma, or leukemia. In someembodiments, the cancer is renal cell carcinoma, malignant melanoma,non-small cell lung cancer (NSCLC), ovarian cancer, Hodgkin's lymphomaor squamous cell carcinoma. In some embodiments, the cancer is selectedfrom breast cancer, colon cancer, rectal cancer, lung cancer, prostatecancer, melanoma, leukemia, ovarian cancer, gastric cancer, renal cellcarcinoma, liver cancer, pancreatic cancer, lymphomas and myeloma. Insome embodiments, the cancer is a solid tumor or hematological cancer.

In some embodiments, the cancer does not have any cells expressing PD-1,PD-L1, or PD-L2 at detectable levels.

In some embodiments, the combination of a compound of Formula (I) andPD-1 inhibitor (or PD-L1 inhibitor/PD-L2 inhibitor) exhibits bettersafety profile and lower toxicity than the combination of CTL.-4 andPD-1 inhibitor (or PD-L1 inhibitor/PD-L2 inhibitor), In someembodiments, the therapeutic index for the combination of a compound ofFormula (I) and PD-1 inhibitor (or PD-L1 inhibitor/PD-L2 inhibitor)greater than the therapeutic index of the combination of CTLA-4 and PD-1inhibitor (or PD-Ll inhibitor/PD-L2 inhibitor). In some embodiments, thecompound of Formula (I) is tucaresol.

Some embodiments relate to a method of disrupting cancer associatedtumor vasculature in a subject comprising co-administering to thesubject a compound of Formula (J) described herein and plinabulin. Insome embodiments, the method of disrupting cancer associated tumorvasculature further includes administering one or more immune checkpointinhibitor. In some embodiments, the compound of Formula (I) istucaresol.

Various cancers are associated the formation of tumor vasculature. Insome embodiments, the cancer is the cancer is selected from the groupconsisting of a melanoma, a pancreatic cancer, a colorectaladenocarcinoma, a brain tumor, acute lymphoblastic leukemia, chroniclymphocytic leukemia, hormone refractory metastatic prostate cancer,metastatic breast cancer, non-small cell lung cancer, renal cellcarcinoma, head and neck cancer, prostate cancer, colon cancer,anaplastic thyroid cancer.

Some embodiments include co-administering a composition, and/orpharmaceutical composition described herein, with an additionalmedicament. For example, as described above, some embodiments includeco-administering a compound of Formula (I) described herein with one ormore immune checkpoint inhibitor, some embodiments includeco-administering a compound of Formula (I) described herein with one ormore immune checkpoint inhibitor and plina.bulin, and some embodimentsinclude co-administering a compound of Formula (I) described herein withplinabulin. In some embodiments, the compound of Formula (I) istucaresol, By “co-administration,” it is meant that the two or moreagents are administered in such a manner that administration of one ormore agent has an effect on the efficacy and/or safety of the one ormore other agent, regardless of when or how they are actuallyadministered. In one embodiment, the agents are administeredsimultaneously. In one such embodiment, administration in combination isaccomplished by combining the agents in a single dosage form, In anotherembodiment, the agents are administered sequentially. In one embodimentthe agents are administered through the same route, such as orally orintravenously. In another embodiment, the agents are administeredthrough different routes, such as one being administered orally andanother being administered i.v. In some embodiments; the time periodbetween administration of one or more agent and administration of theco-administered one or more agent can be about 5 min, 10 min, 20 min, 30min, 40 min, 50 min, 1 hour, 2 hours, 3 hours, 5 hours, 8 hours, 10hours, 12 hours, 15 hours, 18 hours, 20 hours, 24 hours, 36 hours, 48hours, 3 days, 4 days, 5 days, 6 days, 7 days, 10 days, 14 days, 21days, 28 days, or 30 days. In sonic embodiments, the time period betweenadministration of one or more agent and administration of theco-administered one or more agent can be in the range of about 1 min-5min, 1 min-10 min, 1 min-20 min, 1 min-30 min, 1 min-40 min, 1 min-50min, 1 min-1 h, 1 min-2 h, 1 min-4 h, 1 min-6 h, 1 min-8 h, 1 min-10 h,1 min-12 h, 1 min-36 h, 1 min-48 h, 1 min-60 h, 1 min-72 h, 5 min-10min, 5 min-20 min, 5 min-30 min, 5 min-40 min, 5 min-50 min, 5 min-1 h,5 min-2 h, 5 min-4 h, 5 min-6 h, 5 min-8 h, 5 min-10 h, 5 min-12 h, 5min-24 h, 5 min-36 h, 5 min-48 h, 5 min-60 h, 5 min-72 h, 10 min-20 min,10 min-30 min, 10 min-40 min, 10 min-50 min, 10 min-1 h, 10 min-2 h, 10min-4 h, 10 min-6 h, 10 min-8 h, 10 min-10 h, 10 min-12 h, 10 min-24 h,10 min-36 h, 10 min-48 h, 10 min-60 h, 10 min-72 h, 30 min-40 min. 30min-50 min, 30 min-lh, 30 min-2 h, 30 min-4 h, 30 min-6 h, 30 min-8 h,30 min-10 h, 30 min-12 h, 30 min-24 h, 30 min-36 h, 30 min-48 h, 30min-60 h, 30 min-72 h, 1 h-2 h, 1 h-4 h, 1 h-6 h, 1 h-8 h, 1 h-10 h, 1h-12 h, 1 h-24 h, 1 h-36 h, 1 h-48 h, 1 h-60h, 1 h-72 h, 6 h-8 h, 6 h-10h, 6 h-12 h, 6 h-24 h, 6 h-36 h, 6 h-48 h, 6 h-60 h, 6 h-72 h, 12 h-24h, 12 h-36 h, 12 h-48 h, 12 h-60 h, or 12 h-72 h.

The actual dose of the active compounds described herein depends on thespecific compound, and on the condition to be treated; the selection ofthe appropriate dose is well within the knowledge of the skilledartisan. In some embodiments, the method described herein comprisesadministering the compound of Formula (I) at a dose in the range of fromabout 0.01 mg/kg to about 250 mg/kg of body weight, from about 0.1 mg/kgto about 200 mg/kg of body weight, from about 0.25 mg/kg to about 120mg/kg of body weight, from about 0.5 mg/kg to about 70 mg/kg of bodyweight, from about 1.0 mg/kg to about 50 mg/kg of body weight, fromabout 1.0 mg/kg to about 15 mg/kg of body weight, from about 2.0 mg/kgto about 15 mg/kg of body weight, from about 3.0 mg/kg to about 12 mg/kgof body weight, or from about 5.0 mg/kg to about 10 mg/kg of bodyweight. In some embodiments, the method described herein comprisesadministering the compound of Formula (I) at a dose in the range of0.5-1, 0.5-2, 0.5-3, 0.5-4, 0.5-5, 0.5-6, 0.5-7, 0.5-8, 0.5-9, 0.5-10,1-2, 1-3, 1-4, 1-5, 1-6, 1-7, 1-8, 1-9, 1-10, 1-20, 1-30, 1-40, 1-50,1-60, 1-70, 1-80, 1-90, 1-100, 2.5-5, 2.5-10, 2.5-20, 2.5-30, 2.5-40,2.5-50, 2.5-60, 2.5-70, 2.5-80, 2.5-90, 2.5-100, 3-5, 3-10, 3-20, 3-30,3-40, 3-50, 3-60, 3-70, 3-80, 3-90, 3-100, 5-10, 5-20, 5-30, 5-40,5-50,5-60, 5-70, 5-80, 5-90, 5-100, 7.5-10, 7.5-20, 7.5-30, 7.5-40,7.5-50, 7.5-60, 7.5-70, 7.5-80, 7.5-90, 7.5-100, 10-10, 10-20, 10-30,10-40, 10-50, 10-60, 10-70, 10-80, 10-90, 10-100, 10-150, 10-200, 20-30,20-40, 20-50, 20-60, 20-70, 20-80, 20-90, 20-100, 20-150, 20-200, 30-40,30-50, 30-60, 30-70, 30-80, 30-90, 30-100, 30-150, 30-200, 40-50, 40-60,40-70, 40-80, 40-90, 40-100, 40-150, 40-200, 40-300, 50-60, 50-70,50-80, 50-90, 50-100, 50-150, 50-200, 50-250, 50-300, 60-80, 60-100,60-150, 60-200, 70-100, 70-150, 70-200, 70-250, 70-300, 80-100, 80-15080-200, 80-250, 80-300, 90-100, 90-150, 90-200, 90-250, 90-300, 90-350,90-400, 100-150, 100-200, 100-250, 100-300, 100-350, or 100-400 mg/kg ofbody weight. In some embodiments, the compound of Formula (I) describedherein may be administered at a dose of about 0.1, 0.25, 0.5, 2, 3, 4,5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15,16, 17, 18,19, 20, 22.5, 25, 27.5,30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, or 100 mg/kg ofbody weight. In some embodiments, the method described herein comprisesadministering the compound of formula (I) at a dose of about 3 mg/kg. Insome embodiments, the method described herein comprises administeringthe compound of formula (I) at a dose of about 3 mg/kg every three weeksfor a total of four doses. In some embodiments, the compound of Formula(I) is tucaresol.

In some embodiments, the compound of Formula (I) (e.g., tucaresol) isadministered at an amount of about 0.5-1, 0.5-2, 0.5-3, 0.5-4, 0.5-5,0.5-6, 0.5-7, 0.5-8, 0.5-9, 0.5-10, 2.5-3, 2.5-4, 2.5-5, 2.5-6, 2.5-7,2.5-8, 2.5-9, 2.5-10,3-10, 5-10, 1-10, 1-20, 1-30, 1-40, 1-50, 1-60,1-70, 1-80, 1-90, 1-100, 2.5-10, 2.5-20, 2.5-30, 2.5-40, 2.5-50, 2.5-60,2.5-70, 2.5-80, 2.5-90, 2.5-100, 5-10, 5-20, 5-30, 5-40, 5-50,5-60,5-70, 5-80, 5-90, 5-100, 7.5-10, 7.5-20, 7.5-30, 7.5-40, 7.5-50, 7.5-60,7.5-70, 7.5-80, 7.5-90, 7.5-100, 10-10, 10-20, 10-30, 10-40, 10-50,10-60, 10-70, 10-80, 10-90, 10-100, 10-150, 10-200, 20-30, 20-40, 20-50,20-60, 20-70, 20-80, 20-90, 20-100, 20-150, 20-200, 30-40, 30-50, 30-60,30-70, 30-80, 30-90, 30-100, 30-150, 30-200, 40-50, 40-60, 40-70, 40-80,40-90, 40-100, 40-150, 40-200, 40-300, 50-60, 50-70, 50-80, 50-90,50-100, 50-150, 50-200, 50-250, 50-300, 60-80, 60-100, 60-150, 60-200,70-100, 70-150, 70-200, 70-250, 70-300, 70-500, 70-750, 70-1000,70-1500, 70-2000, 70-3000, 80-100, 80-150, 80-200, 80-250, 80-300,80-500, 80-750, 80-1000, 80-1500, 80-2000, 80-3000, 90-100, 90-150,90-200, 90-250, 90-300, 90-350, 90-400, 90-500, 90-750, 90-1000,90-1500, 90-2000, 90-3000, 100-150, 100-200, 100-250, 100-300, 100-350,100-400, 100-500, 100-600, 100-700, 100-800, 100-900, 100-1000,100-1500, 100-2000, 100-2500, 100-3000, 100-3500, 100-4000, 200-500,200-700, 200-1000, 200-1500, 200-2000, 200-2500, 200-3000, 200-3500,200-4000, 500-1000, 500-1500, 500-2000, 500-2500, 500-3000, 500-3500, or500-4000 mg per dose. In some embodiments, the compound of Formula (I)is administered at an amount of about 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7,0.8, 0.9, 1, 1.25, 1.5, 1.75, 2, 2.25, 2.5, 2.75, 3, 3.25, 3.5, 3.75, 4,4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5, 10, 12.5, 15, 17.5, 20,22.5, 25, 27.5, 30, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250, 300,350,400 450, 500, 550, 600, 650, 700, 750, 800, 850, 900, 1000, 1100,1200, 1250, 1300, 1400, 1500, 1600, 1700, 1800, 1900, 2000, 2100, 2200,2300, 2400, 2500, 2600, 2700, 2800, 2900, 3000, 3100, 3200, 3300, 3400,3500, 3600, 3700, 3800, 3900, 4000, 4100, 4200, 4300, 4400, 4500, 4600,4700, 4800, 4900, or 5000 mg per dose. In some embodiments, the compoundof formula (I) is administered at an amount of about 25 mg, 50 mg, or100 mg per dose,

In some embodiments, the method described herein comprises administeringone or more check point inhibitors at a doze in the range of about0.5-1, 0.5-2, 0.5-3, 0.5-4, 0.5-5, 0.5-6, 0.5-7, 0.5-8, 0.5-9, 0.5-10,1-2, 1-3, 1-4, 1-5, 1-6, 1-7, 1-8, 1-9, 1-10, 1-20, 1-30, 1-40, 1-50,1-60, 1-70, 1-80, 1-90, 1-100, 2-3, 2-4, 2-5, 2-6, 2-7, 2-8, 2-9, 2-10,2-20, 2-30, 2-40, 2-50, 2-60, 2-70, 2-80, 2-90, 2-100, 2.5-3, 2.5-3.5,2.5-4, 2.5-5, 2.5-6, 2.5-7, 2.5-9, 2.5-10, 3-4, 3-5, 3-6, 3-7, 3-8, 3-9,3-10, 5-10, 5-20, 5-30, 5-40, 5-50,5-60, 5-70, 5-80, 5-90, 5-100,7.5-10, 7.5-20, 7.5-30, 7.5-40, 7.5-50, 7.5-60, 7.5-70, 7.5-80, 7.5-90,7.5-100, 10-10, 10-20, 10-30, 10-40, 10-50, 10-60, 10-70, 10-80, 10-90,10-100, 10-150, 10-200, 20-30, 20-40, 20-50, 20-60, 20-70, 20-80, 20-90,20-100, 20-150, 20-200, 30-40, 30-50, 30-60, 30-70, 30-80, 30-90,30-100, 30-150, 30-200, 40-50, 40-60, 40-70, 40-80, 40-90, 40-100,40-150, 40-200, 40-300, 50-60, 50-70, 50-80, 50-90, 50-100, 50-150,50-200, 50-250, 50-300, 60-80, 60-100, 60-150, 60-200, 70-100, 70-150,70-200, 70-250, 70-300, 80-100, 80-150, 80-200, 80-250, 80-300, 90-100,90-150, 90-200, 90-250, 90-300, 90-350, 90-400, 100-150, 100-200,100-250, 100-300, 100-350, or 100-400 mg/kg of the body weight. In someembodiments, the method described herein comprised administering one ormore checkpoint inhibitors at a dose of about 0.1, 0.2, 0.3, 0.4, 0.5,0.6, 0.7, 0.8, 0.9, 1, 1.25, 1.5, 1.75, 2, 2.25, 2.5. 2.75, 3, 3.25,3,5, 3.75, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5, 10, 12.5, 15,17.5, 20, 22.5, 25, 27.5, 30, 40, 50, 60, 70, 80, 90 or 100 mg/kg of thebody weight.

In some embodiments, the one or more check point inhibitor areadministered at an amount of about 0.5-1, 0.5-2, 0.5-3, 0.5-4, 0.5-5,0.5-6, 0.5-7, 0.5-8, 0.5-9, 0.5-10, 1-10, 1-20, 1-30, 1-40, 1-50, 1-60,1-70, 1-80, 1-90, 1-100, 1-150, 1-200, 1-250, 1-300, 1-500, 2.5-3,2.5-4, 2.5-5, 2.5-6, 2.5-7, 2.5-8, 2.5-9, 2.5-10, 2.5-20, 2.5-30,2.5-40, 2.5-50, 2.5-60, 2.5-70, 2.5-80, 2.5-90, 2.5-100, 2.5-200,2.5-250, 2.5-300, 2.5-500, 3-10, 3-20, 3-30, 3-40, 3-50, 3-60, 3-70,3-80, 3-90, 3-100, 3-200, 3-250, 3-300, 3-500, 5-10, 5-10, 5-20, 5-30,5-40, 5-50,5-60, 5-70, 5-80, 5-90, 5-100, 7.5-10, 7.5-20, 7.5-30,7.5-40, 7.5-50, 7.5-60, 7.5-70, 7.5-80, 7.5-90, 7.5-100, 10-10, 10-20,10-30, 10-40, 10-50, 10-60, 10-70, 10-80, 10-90, 10-100, 10-150, 10-200,20-30, 20-40, 20-50, 20-60, 20-70, 20-80, 20-90, 20-100, 20-150, 20-200,30-40, 30-50, 30-60, 30-70, 30-80, 30-90, 30-100, 30-150, 30-200, 40-50,40-60, 40-70, 40-80, 40-90, 40-100, 40-150, 40-200, 40-300, 50-60,50-70, 50-80, 50-90, 50-100, 50-150, 50-200, 50-250, 50-300, 60-80,60-100, 60-150, 60-200, 70-100, 70-150, 70-200, 70-250, 70-300, 70-500,70-750, 70-1000, 70-1500, 70-2000, 70-3000, 80-100, 80-150, 80-200,80-250, 80-300, 80-500, 80-750, 80-1000, 80-1500, 80-2000, 80-3000,90-100, 90-150, 90-200, 90-250, 90-300, 90-350, 90-400, 90-500, 90-750,90-1000, 90-1500, 90-2000, 90-3000, 100-150, 100-200, 100-250, 100-300,100-350, 100-400, 100-500, 100-600, 100-700 100-800, 100-900, 100-1000,100-1500, 100-2000, 100-2500, 100-3000, 100-3500, 100-4000, 200-500,200-700, 200-1000, 200-1500, 200-2000, 200-2500, 200-3000, 200-3500,200-4000, 500-1000, 500-1500, 500-2000, 500-2500, 500-3000, 500-3500, or500-4000 mg per dose. In some embodiments, the one or more check pointinhibitors are administered at an amount of about 1, 2, 3, 4, 5, 6, 7,8, 9, 10, 12, 14, 16, 18, 20, 22, 25, 27, 30, 35, 40, 45, 50, 55, 60,65, 70, 75, 80, 85, 90, 95, 100, 110, 120, 130, 140, 150, 160, 170, 180,190, 200, 250, 300, 350, 400, 450, 500, 600, 700, 800, 900, 1000, 1100,1200, 1300, 1400, 1500, 1600, 1700, 1800, 1900, or 2000 mg per dose.

In some embodiments, the method described herein comprises administeringan inhibitor of PD-1 at a dose in the range of about 0.5-1, 0.5-2,0.5-3, 0.5-4, 0.5-5, 0.5-6, 0.5-7, 0.5-8, 0.5-9, 0.5-10, 1-2, 1-3, 1-4,1 5, 1-6, 1-7, 1-8, 1-9, 1-10, 1-20, 1-30, 1-40, 1-50, 1-60, 1-70, 1-80,1-90, 1-100, 2-3, 2-4, 2-5, 2-6, 2-7, 2-8, 2-9, 2-10, 2-20, 2-30, 2-40,2-50, 2-60, 2-70, 2-80, 2-90, 2-100, 2.5-3, 2.5-3.5, 2.5-4, 2.5-5,2.5-6, 2.5-7, 2.5-9, 2.5-10, 3-4, 3-5, 3-6, 3-7, 3-8, 3-9, 3-10, 5-10,5-20, 5-30, 5-40, 5-50,5-60, 5-70, 5-80, 5-90, 5-100, 7.5-10, 7.5-20,7.5-30, 7.5-40, 7.5-50, 7.5-60, 7.5-70, 7.5-80, 7.5-90, 7.5-100, 10-10,10-20, 10-30, 10-40, 10-50, 10-60, 10-70, 10-80, 10-90, 10-100, 10-150,10-200, 20-30, 20-40, 20-50, 20-60, 20-70, 20-80, 20-90, 20-100, 20-150,20-200, 30-40, 30-50, 30-60, 30-70, 30-80, 30-90, 30-100, 30-150,30-200, 40-50, 40-60, 40-70, 40-80, 40-90, 40-100, 40-150, 40-200,40-300, 50-60, 50-70, 50-80, 50-90, 50-100, 50-150, 50-200, 50-250,50-300, 60-80, 60-100, 60-150, 60-200, 70-100, 70-150, 70-200, 70-250,70-300, 80-100, 80-150, 80-200, 80-250, 80-300, 90-100, 90-150, 90-200,90-250, 90-300, 90-350, 90-400, 100-150, 100-200, 100-250, 100-300,100-350, or 100-400 mg/kg of the body weight. In some embodiments, themethod described herein comprises administering the inhibitor of PD-1 ata dose of about 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 1.25,1.5, 1.75, 2, 2.25, 2.5, 2.75, 3, 3.25, 3.5, 3.75, 4, 4.5, 5, 5.5, 6,6.5, 7, 7.5, 8, 8.5, 9, 9.5, 10, 12.5, 15, 17.5, 20, 22.5, 25, 27.5, 30,40, 50, 60, 70, 80, 90 or 100 mg/kg of the body weight. In someembodiments, the inhibitor of PD-1 is administered at a dose of about 3mg/kg. In some embodiments, the inhibitor of PD-1 is administered at adose of about 2 mg/kg.

In some embodiments, the PD-1 inhibitor is administered at an amount ofabout 1-10, 1-20, 1-30, 1-40, 1-50, 1-60, 1-70, 1-80, 1-90, 1-100,1-150, 1-200, 1-250, 1-300, 1-500, 2.5-3, 2.5-4, 2.5-5, 2.5-6, 2.5-7,2.5-8, 2.5-9, 2.5-10, 2.5-20, 2.5-30, 2.5-40, 2.5-50, 2.5-60, 2.5-70,2.5-80, 2.5-90, 2.5-100, 2.5-200, 2.5-250, 2.5-300, 2.5-500, 3-10, 3-20,3-30, 3-40, 3-50, 3-60, 3-70, 3-80, 3-90, 3-100, 3-200, 3-250, 3-300,3-500, 5-10, 5-10, 5-20, 5-30, 5-40, 5-50,5-60, 5-70, 5-80, 5-90, 5-100,7.5-10, 7.5-20, 7.5-30, 7.5-40, 7.5-50, 7.5-60, 7.5-70, 7.5-80, 7.5-90,7.5-100, 10-10, 10-20, 10-30, 10-40, 10-50, 10-60, 10-70, 10-80, 10-90,10-100, 10-150, 10-200, 20-30, 20-40, 20-50, 20-60, 20-70, 20-80, 20-90,20-100, 20-150, 20-200, 30-40, 30-50, 30-60, 30-70, 30-80, 30-90,30-100, 30-150, 30-200, 40-50, 40-60, 40-70, 40-80, 40-90, 40-100,40-150, 40-200, 40-300, 50-60, 50-70, 50-80, 50-90, 50-100 50-150,50-200, 50-250, 50-300, 60-80, 60-100, 60-150, 60-200, 70-100, 70-150,70-200, 70-250, 70-300, 70-500, 70-750, 70-1000, 70-1500, 70-2000,70-3000, 80-100, 80-150, 80-200, 80-250, 80-300, 80-500, 80-750,80-1000, 80-1500, 80-2000, 80-3000, 90-100, 90-150, 90-200, 90-250,90-300, 90-350, 90-400, 90-500, 90-750, 90-1000, 90-1500, 90-2000,90-3000, 100-150, 100-200, 100-250, 100-300, 100-350, 100-400, 100-500,100-600, 100-700, 100-800, 100-900, 100-1000, 100-1500, 100-2000,100-2500, 100-3000, 100-3500, 100-4000, 200-500, 200-700, 200-1000,200-1500, 200-2000, 200-2500, 200-3000, 200-3500, 200-4000, 500-1000,500-1500, 500-2000, 500-2500, 500-3000, 500-3500, or 500-4000 mg perdose. In some embodiments, the PD-1 inhibitor is administered at anamount of about 10-30, 10-50, 10-80, 10-100, 10-125, 10-150, 10-175,10-200, 10-250, 10-300, 10-400, 20-50, 20-100, 20-125, 20-150, 20-175,20-200, 20-250, 20-300, 20-400, 30-50, 30-80, 30-100, 30-125, 30-150,30-175, 30-200, 30-250, 30-300, 30-400, 40-50, 40-80, 40-100, 40-125,40-150, 40-175, 40-200, 40-250, 40-300, 40-400, 50-80, 50-100, 50-125,50-150, 50-175, 50-200, 50-250, 50-300, or 50-400 mg per dose.

In some embodiments, the method described herein comprises administeringan inhibitor of PD-Ll. at a dose in the range of about 0.5-1, 0.5-2,0.5-3, 0.5-4, 0.5-5, 0.5-6, 0.5-7, 0.5-8, 0.5-9, 0.5-10, 1-2, 1-3, 1-4,1-5, 1-6, 1-7, 1-8, 1-9, 1-10, 1-20, 1-30, 1-40, 1-50, 1-60, 1-70, 1-80,1-90, 1-100, 2-3, 2-4, 2-5, 2-6, 2-7, 2-8, 2-9, 2-10, 2-20, 2-30, 2-40,2-50, 2-60, 2-70, 2-80, 2-90, 2-100, 2.5-3, 2.5-3.5, 2.5-4, 2.5-5,2.5-6, 2.5-7, 2.5-9, 2.5-10, 3-4, 3-5, 3-6, 3-7, 3-8, 3-9, 3-10, 5-10,5-20, 5-30, 5-40, 5-50,5-60, 5-70, 5-80, 5-90, 5-100, 7.5-10, 7.5-20,7.5-30, 7.5-40, 7.5-50, 7.5-60, 7.5-70, 7.5-80, 7.5-90, 7.5-100, 10-10,10-20, 10-30, 10-40, 10-50, 10-60, 10-70, 10-80, 10-90, 10-100, 10-150,10-200, 20-30, 20-40, 20-50, 20-60, 20-70, 20-80, 20-90, 20-100, 20-150,20-200, 30-40, 30-50, 30-60, 30-70, 30-80, 30-90, 30-100, 30-150,30-200, 40-50, 40-60, 40-70, 40-80, 40-90, 40-100, 40-150, 40-200,40-300, 50-60, 50-70, 50-80, 50-90, 50-100, 50-150, 50-200, 50-250,50-300, 60-80, 60-100, 60-150, 60-200, 70-100, 70-150, 70-200, 70-250,70-300, 80-100, 80-150, 80-200, 80-250, 80-300, 90-100, 90-150, 90-200,90-250, 90-300, 90-350, 90-400, 100-150, 100-200, 100-250, 100-300,100-350, or 100-400 mg/kg of the body weight. In some embodiments, themethod described herein comprises administering the inhibitor of PD-L1at a dose of about 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0,9, 1, 1.25,1.5, 1.75, 2, 2.25, 2.5, 2.75, 3, 3,25, 3,5, 3.75, 4, 4.5, 5, 5.5, 6,6.5, 7, 7.5, 8, 8.5, 9, 9.5, 10, 12.5, 15, 17.5, 20, 22.5, 25, 27.5, 30,40, 50, 60, 70, 80, 90 or 100 mg/kg of the body weight.

In some embodiments, the PD-L1 inhibitor (e.g., atezolizumab) isadministered at an amount of about 1-10, 1-20, 1-30, 1-40, 1-50, 1-60,1-70, 1-80, 1-90, 1-100, 1-150, 1-200, 1-250, 1-300, 1-500, 2.5-3,2.5-4, 2.5-5, 2.5-6, 2.5-7, 2.5-8, 2.5-9, 2.5-10, 2.5-20, 2.5-30,2.5-40, 2.5-50, 2.5-60, 2.5-70, 2.5-80, 2.5-90, 2.5-100, 2.5-200,2.5-250, 2.5-300, 2.5-500, 3-10, 3-20, 3-30, 3-40, 3-50, 3-60, 3-70,3-80, 3-90, 3-100, 3-200, 3-250, 3-300, 3-500, 5-10, 5-10, 5-20, 5-30,5-40, 5-50,5-60, 5-70, 5-80, 5-90, 5-100, 7.5-10, 7.5-20, 7.5-30,7.5-40, 7.5-50, 7.5-60, 7.5-70, 7.5-80, 7.5-90, 7.5-100, 10-10, 10-20,10-30, 10-40, 10-50, 10-60, 10-70, 10-80, 10-90, 10-100, 10-150, 10-200,20-30, 20-40, 20-50, 20-60, 20-70, 20-80, 20-90, 20-100, 20-150, 20-200,30-40, 30-50, 30-60, 30-70, 30-80, 30-90, 30-100, 30-150, 30-200, 40-50,40-60, 40-70, 40-80, 40-90, 40-100, 40-150, 40-200, 40-300, 50-60,50-70, 50-80, 50-90, 50-100, 50-150, 50-200, 50-250, 50-300, 60-80,60-100, 60-150, 60-200, 70-100, 70-150, 70-200, 70-250, 70-300, 70-500,70-750, 70-1000, 70-1500, 70-2000, 70-3000, 80-100, 80-150, 80-200,80-250, 80-300, 80-500, 80-750, 80-1000, 80-1500, 80-2000, 80-3000,90-100, 90-150, 90-200, 90-250, 90-300, 90-350, 90-400, 90-500, 90-750,90-1000, 90-1500, 90-2000, 90-3000, 100-150, 100-200, 100-250, 100-300,100-350, 100-400, 100-500, 100-600, 100-700, 100-800, 100-900, 100-1000,100-1500, 100-2000, 100-2500, 100-3000, 100-3500, 100-4000, 200-500,200-700, 200-1000, 200-1500, 200-2000, 200-2500, 200-3000, 200-3500,200-4000, 500-1000, 500-1500, 500-2000, 500-2500, 500-3000, 500-3500, or500-4000 mg per dose. In some embodiments, the PD-L1 inhibitor isadministered at an amount of about 500-1500, 600-1500, 700-1500,800-1500, 900-1500, 1000-1500, or 1100-1300 mg per dose. In someembodiments, the PD-L1 inhibitor is administered at an amount of about1200 mg per dose.

In some embodiments, the method described herein comprises administeringthe inhibitor of CTLA-4 (e.g., ipilimumab) at a dose in the range ofabout 0.5-1, 0.5-2, 0.5-3, 0.5-4, 0.5-5, 0.5-6, 0.5-7, 0.5-8, 0.5-9,0.5-10, 1-2, 1-3, 1-4, 1-5, 1-6, 1-7, 1-8, 1-9, 1-10, 1-20, 1-30, 1-40,1-50, 1-60, 1-70, 1-80, 1-90, 1-100, 2-3, 2-4, 2-5, 2-6, 2-7, 2-8, 2-9,2-10, 2-20, 2-30, 2-40, 2-50, 2-60, 2-70, 2-80, 2-90, 2-100, 2.5-3,2.5-3.5, 2.5-4, 2.5-5, 2.5-6, 2.5-7, 2.5-9, 2.5-10, 3-4, 3-5, 3-6, 3-8,3-9, 3-10, 5-10, 5-20, 5-30, 5-40, 5-50,5-60, 5-70, 5-80, 5-90, 5-100,7.5-10, 7.5-20, 7.5-30, 7.5-40, 7.5-50, 7.5-60, 7.5-70, 7.5-80, 7.5-90,7.5-100, 10-10, 10-20, 10-30, 10-40, 10-50, 10-60, 10-70, 10-80, 10-90,10-100, 10-150, 10-200, 20-30, 20-40, 20-50, 20-60, 20-70, 20-80, 20-90,20-100, 20-150, 20-200, 30-40, 30-50, 30-60, 30-70, 30-80, 30-90,30-100, 30-150, 30-200, 40-50, 40-60, 40-70, 40-80, 40-90, 40-100,40-150, 40-200, 40-300, 50-60, 50-70, 50-80, 50-90, 50-100, 50-150,50-200, 50-250, 50-300 mg/kg of the body weight. In some embodiments,the method described herein comptises administering the inhibitor ofCTLA-4 at a dose of about 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9,1, 1.25, 1.5, 1.75, 2, 2.25, 2.5, 2.75, 3, 3.25, 3.5, 3.75, 4, 4.5, 5,5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5, 10, 12.5, 15, 17.5, 20, 22.5, 25,27.5, 30, 40, 50, 60, 70, 80, 90 or 100 mg/kg of the body weight. Insome embodiments, the inhibitor of CTLA-4 is administered at a dose ofabout 3 mg/kg. In some embodiments, the inhibitor of CTLA-4 isadministered at a dose of lower than 3 mg/kg. In some embodiments, theinhibitor of CTLA-4 is administered at a dose of about 0.5, 1, 1.5, 2,or 2.5 mg/kg.

In some embodiments, the CTLA-4 inhibitor is administered at an amountof about 1-10, 1-20, 1-30, 1-40, 1-50, 1-60, 1-70, 1-80, 1-90, 1-100,1-150, 1-200, 1-250, 1-300, 1-500, 2.5-3, 2.5-4, 2.5-5, 2.5-6, 2.5-7,2.5-8, 2.5-9, 2.5-10, 2.5-20, 2.5-30, 2.5-40, 2.5-50, 2.5-60, 2.5-70,2.5-80, 2.5-90, 2.5-100, 2.5-200, 2.5-250, 2.5-300, 2.5-500, 3-10, 3-20,3-30, 3-40, 3-50, 3-60, 3-70, 3-80, 3-90, 3-100, 3-200, 3-250, 3-300,3-500, 5-10, 5-10, 5-20, 5-30, 5-40, 5-50,5-60, 5-70, 5-80, 5-90, 5-100,7.5-10, 7.5-20, 7.5-30, 7.5-40, 7.5-50, 7.5-60, 7.5-70, 7.5-80, 7.5-90,7.5-100, 10-10, 10-20, 10-30, 10-40, 10-50, 10-60, 10-70, 10-80, 10-90,10-100, 10-150, 10-200, 20-30, 20-40, 20-50, 20-60, 20-70, 20-80, 20-90,20-100, 20-150, 20-200, 30-40, 30-50, 30-60, 30-70, 30-80, 30-90,30-100, 30-150, 30-200, 40-50, 40-60, 40-70, 40-80, 40-90, 40-100,40-150, 40-200, 40-300, 50-60, 50-70, 50-80, 50-90, 50-100, 50-150,50-200, 50-250, 50-300, 60-80, 60-100, 60-150, 60-200, 70-100, 70-150,70-200, 70-250, 70-300, 70-500, 70-750, 70-1000, 70-1500, 70-2000,70-3000, 80-100, 80-150, 80-200, 80-250, 80-300, 80-500, 80-750,80-1000, 80-1500, 80-2000, 80-3000, 90-100, 90-150, 90-200, 90-250,90-300, 90-350, 90-400, 90-500, 90-750, 90-1000, 90-1500, 90-2000,90-3000, 100-150, 100-200, 100-250, 100-300, 100-350, 100-400, 100-500,100-600, 100-700 100-800, 100-900, 100-1000, 100-1500, 100-2000,100-2500, 100-3000, 100-3500, 100-4000, 200-500, 200-700, 200-1000,200-1500, 200-2000, 200-2500, 200-3000, 200-3500, 200-4000, 500-1000,500-1500, 500-2000, 500-2500, 500-3000, 500-3500, or 500-4000 mg perdose. In some embodiments, the CTLA-4 inhibitor is administered at anamount of about 10-30, 10-50, 10-80, 10-100, 10-125, 10-150, 10-175,10-200, 10-250, 10-300, 10-400, 20-50 20-100, 20-125, 20-150, 20-175,20-200, 20-250, 20-300, 20-400, 30-50, 30-80, 30-100, 30-125, 30-150,30-175, 30-200, 30-250, 30-300, 30-400, 40-50, 40-80, 40-100, 40-125,40-150, 40-175, 40-200, 40-250, 40-300, 40-400, 50-80, 50-100, 50-125,50-150, 50-175, 50-200, 50-250, 50-300, or 50-400 mg per dose,

In some embodiments, the treatment schedule includes co-administrationof the compound of formula (I) (e,g., tucaresol) and one or morecheckpoint inhibitors (e,g., PD-1/PD-L1 inhibitor and CTLA-4 inhibitor)once every 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks,or 8 weeks. In some embodiments, the treatment schedule includesco-administration of the compound of formula (J) and one or morecheckpoint inhibitors once every 2 weeks or 3 weeks. In someembodiments, the treatment schedule includes co-administration of thecompound of formula (I) and one or more checkpoint inhibitors two timesevery 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, or 8weeks. In some embodiments, the treatment schedule includesco-administration of a chemotherapeutic agent and plinabulin once every1 week in a treatment cycle of 1 week, 2 weeks, 3 weeks, 4 weeks, 5weeks, 6 weeks, 7 weeks, or 8 weeks. In some embodiments, the treatmentschedule includes co-administration of the compound of formula (I) andone or more checkpoint inhibitors twice every 1 week in a treatmentcycle of I week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks,or 8 weeks. In some embodiments, the treatment schedule includesco-administration of the compound of formula (I) and one or morecheckpoint inhibitors three times every 1 week, 2 weeks, 3 weeks, 4weeks, 5 weeks, 6 weeks, 7 weeks, or 8 weeks. In some embodiments, thetreatment schedule includes co-administration of the compound of formula(I) and one or more checkpoint inhibitors four times every 1 week, 2weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, or 8 weeks, In someembodiments, the treatment schedule includes co-administration of thecompound of formula (I) and one or more checkpoint inhibitors five timesevery 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, or 8weeks. In some embodiments, the treatment schedule includesco-administration of the compound of formula (I) and one or morecheckpoint inhibitors six times every 1 week, 2 weeks, 3 weeks, 4 weeks,5 weeks, 6 weeks, 7 weeks, or 8 weeks. In some embodiments, thetreatment schedule includes co-administration of the compound of formula(I) and one or more checkpoint inhibitors daily every 1 week, 2 weeks, 3weeks, or 4 weeks. In some embodiments, co-administration of thecompound of formula (I) and one or more checkpoint inhibitors includesadministering the compound of formula (I) prior to administering the oneor more checkpoint inhibitors. In some embodiments, the treatmentschedule includes co-administration of the compound of formula (I) andone or more checkpoint inhibitors 1, 2, 3, 4, 5, 6, or 7 times per day.In some embodiments, the treatment schedule includes co-administrationof the compound of formula (I) and one or more checkpoint inhibitorsonce every 2, 3, 4, 5, or 6 days.

In some embodiments, co-administration of the compound of formula (I)and one or more checkpoint inhibitors includes administering thecompound of formula (I) after administering the one or more checkpointinhibitors. In some embodiments, co-administration of the compound offormula (I) and one or more checkpoint inhibitors includes administeringthe compound of formula (I) concurrently with the one or more checkpointinhibitors. When more than one checkpoint inhibitors are administered,the two check point inhibitors can be administered separately orconcurrently.

In some embodiments, when the compound of formula (I) is administeredprior to the one or more checkpoint inhibitors are administered, the oneor more checkpoint inhibitors can be administered about 1 min, 5 min, 10min, 15 min, 20 min, 25 min, 30 min, 1 h, 1.5 h, 2 h, 2.5 h, 3 h, 4 h, 5h, 6 h, 7 h, 8 h, 9 h, 10 h, 11 h, or 12 h after the administration ofthe compound of formula (I). In some embodiments, the one or morecheckpoint inhibitors are administered in less than about 1 min, 5 min,10 min, 15 min, 20 min, 25 min, 30 min, 1 h, 1.5 h, 2 h, 2.5 h, 3 h, 4h, 5 h, 6 h, 7 h, 8 h, 9 h, 10 h, 1 th, 12 h, 13 h, 14 h, 15 h, 16 h, 17h, 18 h, 19 h, 20 h, 21 h, 22 h, 23 h, or 24 h after the administrationof the compound of formula (I). In some embodiments, the one or morecheckpoint inhibitors are administered in more than about 1 min, 5 min,10 min, 15 min, 20 min, 25 min, 30 min, th, 1.5h, 2 h, 2.5 h, 3 h, 4 h,5 h, 6 h, 7 h, 8 h, 9 h, 10 h, 11 h, 12 h, 13 h, 14 h, 15 h, 16 h, 17 h,18 h, 19 h, 20 h, 21 h, 22 h, 23 h, or 24 h after the administration ofthe compound of formula (I). In some embodiments, the one or morecheckpoint inhibitors are administered in about 1 min-5 min, 1 min-10min, 1 min-20 min, 1 min-25 min, 1 min-30 min, 0.25 h-0.5 h, 0.25-0.75h, 0.25-1 h,0.5h-1 h, 0.5 h-2 h, 0.5 h-2.5 h, 1 h-2 h, 1 h-3 h, 1 h-5 hafter the administration of the compound of formula (I). In someembodiments, the one or more checkpoint inhibitors are administered inabout 1 min-5 min, 1 min-10 min, 1 min-20 min, 1 min30 min, 1 min-40min, 1 min-50 min, 1 min-1 h, 1 min-2 h, 1 min-4 h, 11. min-6 h, 1 min-8h, 1 min-10 h, h, 1 min-24 h, 1 min-36 h, 1 min-48 h, 1 min-60 h, 1min-72 h, 5 min-10 min, 5 min-20 min, 5 min-30 min, 5 min-40 min, 5min-50 min, 5 min-1 h, 5 min-2 h, 5 min-4 h, 5 min-6 h, 5 min-8 h, 5min-10 h, 5 min-12 h, 5 min-24 h, 5 min-36 h, 5 min-48 h, 5 min-60 h, 5min-72 h, 10 min-20 min, 10 min-30 min, 10 min-40 min, 10 min-50 min, 10min-1 h, 10 min-2 h, 10 min-4 h, 10 min-6 h, 10 min-8 h, 10 min-10 h, 10min-12 h, 10 min-24 h, 10 min-36 h, 10 min-48 h, 10 min-60 h, 10 min-72h, 30 min-40 min, 30 min-50 min, 30 min-1 h, 30 min-2 h, 30 min-4 h, 30min-6 h, 30 min-8 h, 30 min-10 h, 30 min-12 h, 30 min-24 h, 30 min-36 h,30 min-48 h, 30 min-60 h, 30 min-72 h, 1 h-2 h, 1 h-4 h, 1 h-6 h, 111-8h, 1 h-10 h, 1 h-12 h, 1 h-24 h, 1 h-36 h, 1 h-48 h, 1 h-60 h, 1 h-72 h,6 h-8 h, 6 h-10 h, 6 h-12. h, 6 h-24 h, 6 h-36 h, 6 h-48 h, 6 h-60 h, 6h-72 h, 12 h-24 h, 12 h-36 h, 12 h-48 h, 12 h-60 h, or 12 h-72 h afterthe administration of the compound of formula (I).

In some embodiments, when the one or more checkpoint inhibitors areadministered prior to the compound of formula (I) is administered, theone or more checkpoint inhibitors are administered about 1 min-5 min, 1min-10 min, 1 min-15 min, 1 min-20 min, 1 min-25 min, 1 min-30 min, 0.25h-0.5 h, 0.25-0.75 h, 0.25-1 h,0.5 h-1 h, 0.5 h-2 h, 0.5 h-2.5 h, 1 h-2h, 1 h-3 h, or 1 h-5 h before the administration of the compound offormula (I). In some embodiments, the one or more checkpoint inhibitorsare administered about 1 min, 5 min, 10 min, 15 min, 20 min, 25 min, 30min, 1 h, 1.5 h, 2 h, 2.5 h, 3 h, 4 h, 5 h, 6 h, 7 h, 8 h, 9 h, 10 h, 11h, or 12 h before the administration of the compound of formula (I). Insome embodiments, the one or more checkpoint inhibitors are administeredless than about 1 min, 5 min, 10 min, 15 min, 20 min, 25 min, 30 min, 1h, 1.5 h, 2 h, 2.5 h, 3 h, 4 h, 5 h, 6 h, 7 h, 8 h, 9 h, 10 h, 11 h, 12h, 13 h, 14 h, 15 h, 16 h, 17 h, 18 h, 19 h, 20 h, 21 h, 22 h, 23 h, or24 h before the administration of the compound of formula (I). In someembodiments, the one or more checkpoint inhibitors are administered morethan about 1 min, 5 min, 10 min, 15 min, 20 min, 25 min, 30 min, 1 h,1.5 h, 2 h, 2.5 h, 3 h, 4 h, 5 h, Oh, 7 h, 8 h, 9 h, 10 h, 11 h, 12 h,13 h, 14 h, 15 h, 16 h, 17 h, 18 h, 19 h, 20 h, 21 h, 22 h, 23 h, or 24h before the administration of the compound of formula (I). In someembodiments, the one or more checkpoint inhibitors are administered inabout 1 min-5 min, 1 min-10 min, 1 min-20 min, 1 min-30 min, 1 min-40min, 1 min-50 min, 1 min-1 h, 1 min-2 h, 1 min-4 h, 1 min-6 h, 1 min-8h, 1 min-10 h, 1 min-12 h, 1 min-24 h, 1 min-36 h, 1 min-48 h, 1 min-60h, 1 min-72 h, 5 min-10 min, 5 min-20 min, 5 min-30 min, 5 min-40 min, 5min-50 min, 5 min-1 h, 5 min-2 h, 5 min-4 h, 5 min-6 h, 5 min-8 h, 5min-10 h, 5 min-12 h, 5 min-24 h, 5 min-36 h, 5 min-48 h, 5 min-60 h, 5min-72 h, 10 min-20 min, 10 min-30 min, 10 min-40 min, 10 min-50 min, 10min-1 h, 10 min-2 h, 10 min-4 h, 10 min-6 h, 10 min-8 h, 10 min-10 h, 10min-12 h, 10 min-24 h, 10 min-36 h, 10 min-48 h, 10 min-60 h, 10 min-72h, 30 min-40 min, 30 min-50 min, 30 min-1 h, 30 min-2 h, 30 min-4 h, 30min-6 h, 30 min-8 h, 30 min-10 h, 30 min-12 h, 30 min-24 h, 30 min-36 h,30 min-48 h, 30 min-60 h, 30 min-72 h, 1 h-2 h, 1 h-4 h, 1 h-6 h, 1 h-8h, 1 h-10 h, 1 h-12 h, 1 h-24 h, 1 h-36 h, 1 h-48 h, 1 h-60 h, 1 h-72 h,6 h-8 h, 6 h-10 h, 6 h-12 h, 6 h-24 h, 6 h-36 h, 6 h-48 h, 6 h-60 h, 6h-72 h, 12 h-24 h, 12 h-36 h, 12 h-48 h, 12 h-60 h, or 12 h-72 h beforethe administration of the compound of formula (I).

The treatment cycle can be repeated as long as the regimen is clinicallytolerated. In some embodiments, the treatment cycle for the compound offormula (I) and the one or more checkpoint inhibitors is repeated for ntimes, wherein n is an integer in the range of 2 to 30. In someembodiments, n is 2, 3, 4, 5, 6, 7, 8, 9, or 10. In some embodiments, anew treatment cycle can occur immediately after the completion of theprevious treatment cycle. In some embodiments, a washout period canoccur before starting a new treatment cycle. In some embodiments, thewashout period can be 1 week, 2 weeks, 3 weeks, or 4 weeks. In someembodiments, the dose of the compound of formula (I) can be the same foreach treatment cycle. In some embodiments, the dose of the compound offormula (I) can be different in each treatment cycle (e.g., the dose canbe 20 mg for the first treatment cycle, 50 mg for the second treatmentcycle, 100 mg for the third treatment cycle).

In some embodiments, after the compound of formula (I) and the one ormore checkpoint inhibitors are administered in one cycle of treatment,the next treatment cycle may include administering only the compound offormula (I). In some embodiments, after the compound of formula (I) andthe one or more checkpoint inhibitors are administered in one cycle oftreatment, the next treatment cycle may include administering both thecompound of formula (I) and the one or more checkpoint inhibitors.

In some embodiments, the compound of formula (I) (e.g., tucaresol) isadministered at a dose of about 3 mg/kg every three weeks as a treatmentcycle and the treatment cycle is repeated four times. In someembodiments, the one or more checkpoint inhibitors (e.g., any one ofPD-1 inhibitor, PD-L1 inhibitor, CTLA-4 inhibitor, and any combinationsthereof) can be co-administered with the compound of formula (I) in eachtreatment cycle. In some embodiments, the one or more checkpointinhibitors can be co-administered with the compound of formula (I) inhalf of the treatment cycles (e.g. the first and the third treatmentcycles).

In some embodiments, the method described herein can include one or moreadditional medicaments. Examples of additional medicaments include otherchemotherapeutic agents.

In some embodiments, the chemotherapeutic agent can be selected from thegroup consisting of Abiraterone Acetate, Abitrexate (Methotrexate),Abraxane (Paclitaxel Albumin-stabilized Nanoparticle Formulation), ABVD,AVE, ABVE-PC, AC, AC-T, Adcetris (Brentuximab Vedotin), ADE,Ado-Trastuzumab Etntansine, Adriamycin (Doxorubicin Hydrochloride),Afatinib Dimaleate, Afinitor (Everolimus), Akynzeo (Netupitant andPalonosetron Hydrochloride), Aldara (Imiquimod), Aldesleukin, Alecensa(Alectinib), Alectinib, Alemtuzumab, Alimta (Pemetrexed Disodium), Aloxi(Palonosetron Hydrochloride), Ambochlorin (Chlorambucil), Amboclorin(Chlorambucil), Aminolevulinic Acid, \Anastrozole, Aprepitant, Aredia(Pamidronate Disodium), Arimidex (Anastrozole), Aromasin (Exemestane),Arranon (Nelarabine), Arsenic Trioxide, Arzerra (Ofatumumab),Asparaginase Erwinia chrysanthemi, Avastin (Bevacizumab)Axitinib,Azacitidine, BEACOPP, Becenum (Carmustine), Beleodaq (Belinostat),Belinostat, Bendamustine Hydrochloride, BEP, Bevacizumab, Bexa.rotene,Bexxar (Tositumomab and Iodine I 131 Tositumomab), Bicalutamide, BiCNU(Carmustine), Bleomycin, Blinatumomab, Blincyto (Blinatumomab),Bortezomib, Bosulif (Bosutinib), Bosutinib, Brentuximab Vedotin,Busulfan, Cabazitaxel, Cabozantinib-S-Malate, CAF, Campath(Alemtuzumab), Camptosar (Irinotecan Hydrochloride), Capecitabine,CAPDX, Carac (Fluorouracil—Topical), Carboplatin, CARBOPLATIN-TAXOL,Carfilzomib, Carmubris (Carmustine), Carmustine, Carmustine implant,Casodex (Bicalutamide), CeeNU (Lomustine), Ceritinib, Cerubidine(Daunorubicin Hydrochloride), Cervarix (Recombinant HPV BivalentVaccine), Cetuxitnab, Chlorambucil,CHLORAMBUCIL-PREDNISONE, CHOP,Cisplatin, Clafen (Cyclophosphamide), Clofarabine, Clofa.rex(Clofarabine), Clolar (Clofarabine), CMF, Cobimetinib, Cometriq(Cabozantinib-S-Malate), COPDAC, COPP, COPP-ABV, Cosmegen(Dactinomycin), Cotellic (Cobimetinib), Crizotinib, CVP,Cyclophosphamide, Cyfos (Ifosfamide), Cyramza (Ramucirumab), Cytarabine,Cytarabine Liposome, Cytosar-U (Cytarabine), Cytoxan (Cyclophosphamide),Dabrafenib, Dacarbazine, Dacogen (Decitabine), Dactinomycin,Daratumumab, Darzalex (Daratumumab), Dasatinib, DaunorubicinHydrochloride, Decitabine, Degarelix, Denileukin Diftitox, Denosumab,DepoCyt (Cytarabine Liposome), Dexamethasone, Dexrazoxane Hydrochloride,Dinutuximab, Docetaxel, Doxil (Doxorubicin Hydrochloride Liposome),Doxorubicin Hydrochloride, Doxorubicin Hydrochloride Liposome, Dox-SL(Doxorubicin Hydrochloride Liposome), DTIC-Dome (Dacarbazine), Efudex(Fluorouracil—Topical), Elitek (Rasburicase), Ellence (EpirubicinHydrochloride), El otuzumab, El oxatin (Oxaliplatin), EltrombopagOlamine, Emend (Aprepitant), (Elotuzumab), Enzalutamide, :EpirubicinHydrochloride, EPOCH, Erbitux (Cetuximab), Eribulin Mesylate, Erivedge(Vismodegib), Erlotinib Hydrochloride, Erwinaze (Asparaginase Erwiniachrysanthemi), Etopophos (Etoposide Phosphate), Etoposide, EtoposidePhosphate, Evacet (Doxorubicin Hydrochloride Liposome), Everolimus,Evista (Raloxifene Hydrochloride), Exemestane, 5-FU (FluorouracilInjection), 5-FU (Fluorouracil—Topical), Fareston (Toremifene), Farydak(Panobinostat), Faslodex (Fulvestrant), FEC, Femara (Letrozole),Filgrastim, Fludara (Fludarabine Phosphate), Fludarabine Phosphate,Fluoroplex (Fluorouracil—Topical), Fluorouracil Injection,Fluorouracil—Topical, Flutamide, Fol ex (Methotrexate), Folex PFS(Methotrexate), FOLFIRI, FOLFTRI-BEVACIZUMAB, FOLFIRI-CETUXIMAB,FOLFIRINOX, FOLFOX, Folotyn (Pralatrexate), FU-LV, Fulvestrant, Gardasil(Recombinant HPV Quadrivalent Vaccine), Gardasil 9 (Recombinant HPVNonavalent Vaccine), Gazyva (Obinutuzumab), Gefitinib, GemcitabineHydrochloride, GEMCITABINE-CISPLATIN, GEMCITABINE-OXALIPLATIN,Getntuzumab Ozogamicin, Gemzar (Gemcitabine Hydrochloride), Gilotrif(Afatinib Dimaleate), Gleevec (Imatinib Mesylate), Gliadel (CarmustineImplant), Gliadel wafer (Carmustine Implant), Glucarpidase, GoserelinAcetate, Halaven (Eribullin Mesylate),Herceptin (Trastuzumab), HPVBivalent Vaccine, Recombinant, HPV Nonavalem Vaccine, Recombinant, HPVQuadrivalent Vaccine, Recombinant, Hycamtin (Topotecan Hydrochloride),Hyper-CVAD, Ibrance (Palbociclib), Ibritumomab Tiuxetan, Ibrutinib, ICE,Iclusig (Ponatinib Hydrochloride), Idamycin (Idarubicin Hydrochloride),Idelalisib, Ifex (Ifosfamide), Ifosfamide, IL-2 (Aldesleukin), ImatinibMesylate, Imbruvica Imiquitnod, Imlygic (Talimogene Laherparepvec),Inlyta (Axitinib), Interferon Alfa-2b, Recombinant, Interleukin-2(Aldesleukin), Intron A (Recombinant Interferon Alfa-2b), Iodine I 131Tositumomab and Tositumomab, Ipilimumab, tressa (Gefitinib), IrinotecanHydrochloride, Irinotecan Hydrochloride Liposome, Istodax (Romidepsin),Ixabepilone, Ixazomib Citrate, Ixempra (Ixabepilone), Jakafi(Ruxolitinib Phosphate), Jevtana (Cabazitaxel), Kadcyla (Ado-TrastuzumabEmtansine), Keoxifene (Raloxifene Hydrochloride), Kepivance(Palifermin), Keytruda (Pembrolizutnab), Kyprolis (Carfilzomib),Lanreotide Acetate, Lapatinib Ditosylate, Lenalidomide, LenvatinibMesylate, (Lenvatinib Mesylate), Letrozole, Leucovorin Calcium, Leukeran(Chlorambucil), Leuprolide Acetate, Levulan (Aminolevulinic Acid),Linfolizin (Chlorambucil), LipoDox (Doxorubicin Hydrochloride Liposome),Lomustine, Lonsurf (Trifluridine and Tipiracil Hydrochloride), Lupron(Leuprolide Acetate), Lupron Depot (Leuprolide Acetate), LupronDepot-Ped (Leuprolide Acetate), Lupron Depot-3 Month (LeuprolideAcetate), Lupron Depot-4 Month (Leuprolide Acetate), Lynparza(Olaparib), Margibo (Vincristine Sulfate Liposome), Matulane(Procarbazine Hydrochloride), Mechlorethamine Hydrochloride, Megace(Megestrol Acetate), Megestrol Acetate, Mekinist (Trametinib),Mercaptopurine, Mesita, Mesnex (Mesna), Methazolastone (Temozolomide),Methotrexate, Methotrexate LIEF (Methotrexate), Mexate (Methotrexate),Mexate-AQ (Methotrexate), Mitomycin C, Mitoxantrone Hydrochloride,IN/litozytrex (Mitomycin C), MOPP,Mozobil (Plerixafor), Mustargen(Mechlorethamine Hydrochloride), Mutamycin (Mitomycin C), Myleran(Busulfan), Mylosar (Azacitidine), Mylotarg (Gemtuzumab Ozogamicin),Nanoparticle Paclitaxel (Paclitaxel Albumin-stabilized NanoparticleFormulation), Navelbine (Vinorelbine Tartrate), Necitumumab, Nelarabine,Neosar (Cyclophosphamide), Netupitant and Palonosetron Hydrochloride,Neupogen (Filgrastim), Nexavar (Sorafenib Tosylate) Nilotinib, Ninlaro(Ixazomib Citrate), Nivolumab, Nolvadex (Tamoxifen Citrate), Nplate(Romiplostim), Obinutuzumab, Odomzo (Sonidegib), OEPA, Ofatumumab, OFF,Olaparib, Omacetaxine Mepesuccinate, Oncaspar (Pegaspargase),Ondansetron Hydrochloride, Onivyde (Irinotecan Hydrochloride Liposome),Ontak (Denileukin Diftitox), Opdivo (Nivolumab), OPPA, Osimertinib,Oxaliplatin, Paclitaxel, Paclitaxel Albumin-stabilized NanoparticleFormulation, PAD, Palbociclib, Palifermin, Palonosetron Hydrochloride,Palonosetron Hydrochloride and Netupitant, Pamidronate Disodium,Panitumumab, Panobinostat, Paraplat (Carboplatin), Paraplatin(Carboplatin), Pazopanib Hydrochloride, PCV, Pegaspargase, PeginterferonAlfa-2b, PEG-Intron (Peginterferon Alfa-2b), Pembrolizumab, PemetrexedDisodium Perieta (Pertuzumab), Pertuzumab, Platinol (Cisplatin),Platinol-AQ (Cisplatin), Plerixafor, Pomalidomide, Pomalyst(Pomalidomide), Ponatinib Hydrochloride, Portrazza (Necitumumab),Pralatrexate, Prednisone, Procarbazine Hydrochloride, Proleukin(Aldesleukin), Prolia (Denosumab), Promacta (Eltrombopag Olamine),Provenge (Sipuleucel-T), Purinethol (Mercaptopurine), Purixan(Mercaptopurine), Radium 223 Dichloride, Raloxifene Hydrochloride,Ramucinunab, Rasburicase, R-CHOP, R-CVP, Recombinant Human.Papillomavirus (HPV) Bivalent Vaccine, Recombinant Human Papillomavirus(HPV) Nonavalent Vaccine, Recombinant Human Papillomavirus (HPV)Quadrivalent Vaccine, Recombinant Interferon Alfa-2b, Regorafenib,R-EPOCH, Revlimid (Lenalidomide), Rheumatrex (Methotrexate), Rituximab,Rolapitant Hydrochloride, Romidepsin, Romiplostim, Rubidomycin(Daunorubicin Hydrochloride), Ruxolitinib Phosphate, SclerosolIntrapleural Aerosol (Talc),Siltuximab, Sipuleucel-T, Somatuline Depot(Lanreotide Acetate), Sonidegib, Sorafenib Tosylate, Sprycel(Dasatinib), STANFORD V, Sterile Talc Powder (Talc), Steritalc (Talc),Stivarga (Regorafenib), Sunitinib Malate, Sutent (Sunitinib Malate),Sylatron (Peginterferon Alfa-2b), Sylvant (Siltuximab), Synovir(Thalidomide), Synribo (Omacetaxine Mepesuccinate), Tabloid(Thioguanine), TAC, Tafinlar (Dabrafenib), Tagrisso (Osimertinib), Talc,Talimogene Laherparepvec, Tamoxifen Citrate, Tarabine PFS (Cytarabine),Tarceva (Erlotinib Hydrochloride), Targretin (Bexarotene), Tasigna(Nilotinib), Taxol (Paclitaxel), Taxotere (Docetaxel), Temodar(Temozolomide), Temozolomide, Temsirolimus, Thalidomide, Thioguanine,Thiotepa, Tolak (Fluorouracil—Topical), Toposar (Etoposide), TopotecanHydrochloride, Toremifene, Torisel (Temsirolimus), Tositumomab andiodine I 131, Tositumomab, Totect (Dexrazoxane Hydrochloride), TPF,Trabectedin, Trametinib, Trastuzumab, Treanda (BendamustineHydrochloride), Trifluridine and Tipiracil Hydrochloride, Trisenox(Arsenic Trioxide), Tykerb (Lapatinib Ditosylate), Unituxin(Dinutuximab), Uridine Triacetate, VAC, Vandetanib, VAMP, Varubi(Rolapitant Hydrochloride), Vectibix (Panitumumab), VeIP, Velban(Vinblastine Sulfate), Velcade (Bortezomib), Velsar (VinblastineSulfate), Vemurafenib, VePesid (Etoposide), Viadur (Leuprolide Acetate),Vidaza (Azacitidine), Vinblastine Sulfate, Vincasar PFS (VincristineSulfate), Vincristine Sulfaw, Vincristine Sulfate Liposome, VinorelbineTartrate, VIP, Vistnodegib, Vistogard (Uridine Triacetate), Voraxaze(Glucarpidase), Vorinostat, Votrient (Pazopanib Hydrochloride),Wellcovorin (Leucovorin Calcium), Xalkori (Crizotinib), Xeloda(Capecitabine), XELIRLXELOX, Xgeva (Denosumab), Xofigo (Radium 223Dichloride), Xtandi (Enzalutamide), Yervoy (Ipilimumab),Yondelis(Trabectedin), Zaltrap (Ziv-Affibercept), Zarxio (Filgrastim), Zelboraf(Vemurafenib), Zevalin (Ibritumomab Tiuxetan), Zinecard (DexrazoxaneHydrochloride), Ziv-Affibercept, Zofran (Ondansetron Hydrochloride),Zoladex (Goserelin Acetate), Zoledronic Acid, Zolinza (Vorinostat),Zorneta (Zoledronic Acid), Zydelig (Idelalisib), Zykadia (Ceritinib),and Zytiga (Abiraterone Acetate)

To further illustrate this invention, the following examples areincluded. The examples should not, of course, be construed asspecifically limiting the invention. Variations of these examples withinthe scope of the claims are within the purview of one skilled in the artand are considered to fall within the scope of the invention asdescribed, and claimed herein. The reader will recognize that theskilled artisan, armed with the present disclosure, and skill in the artis able to prepare and use the invention without exhaustive examples.

EXAMPLES Example 1 Effects on Tumor and Non-Tumor Cell Vaibility 1)Tucaresol

Tucaresol (0-1200 μM) is exposed for 72 hours to a panel of humanliquid, hematological, and solid tumors such as multiple myeloma,leukemia, colorectal, non-small cell lung cancer (squamous andadenocarcinoma), hepatocellular, renal, pancreatic and breast cancercell lines, and human non-tumor such as HUVEC, PBMC, skin fibroblastcells lines. Tucaresol is studied either alone or in combination withstandard-of-care agents (1-100 μM). All cell lines are grown in standardserum-containing media with an exposure time of 24-144 hours. Cellviability is measured using, for example, the Cell TiterGlo® ViabilityAssay. The potency (IC₅₀) and efficacy (% cell kill) are determined fromthe percent cell growth of the vehicle control.

2) Tucaresol Plus PD-1 Antibody

Tucaresol (0-1200 μM) in the presence of a PD-1 antibody is exposed for72 hours to a panel of human liquid, hematological, and solid tumor suchas multiple myeloma, leukemia, colorectal, non-small cell lung cancer(squamous and adenocarcinoma), hepatocellular, renal, pancreatic andbreast cancer cell lines, and human non-tumor such as HUVEC, PBMC, skinfibroblast cells lines, and the viability of the cell lines are measuredas described above. The viability of the cell lines in the presence oftucaresol plus PD-1 antibody is compared to the viability of the celllines in the presence of a CTLA-4 antibody plus the PD-1 antibody orPD-i antibody alone.

3) CTLA-4 Antibody Plus PD-1 Antibody

CTLA-4 antibody in the presence of a PD-1 antibody is exposed for 72hours to a panel of human liquid, hematological, and solid tumor such asmultiple myeloma, leukemia, colorectal, non-small cell lung cancer(squamous and adenocarcinoma), hepatocellular, renal, pancreatic andbreast cancer cell lines, and human non-tumor such as HUVEC, PBMC, skinfibroblast cells lines, and the viability of the cell lines are measuredas described above.

4) Tucaresol Plus Plinabulin

Tucaresol (0-1200 μM) in the presence of Plinabulin is exposed for 72hours to a panel of human liquid, hematological, and solid tumor such asmultiple myeloma, leukemia, colorectal, non-small cell lung cancer(squamous and adenocarcinoma), hepatocellular, renal, pancreatic andbreast cancer cell lines, and human non-tumor such as HUVEC, PBMC, skinfibroblast cells lines, and the viability of the cell lines are measuredas described above.

The viability of the cell lines in the presence of tucaresol, tucaresolplus PD-1 antibody, CTLA-4 antibody plus the PD-1 antibody, andtucaresol plus plinabulin are compared.

Example 2 Potentiation of T Cell Proliferative Responses

Five groups including tucaresol, tucaresol plus PD-1 or PD-L1 antibody,tucaresol plus CTLA-4 antibody, CTLA-4 antibody plus PD-1 or PD-L1antibody, and tucaresol plus plinabulin are tested to determine thepotentiation of T cell proliferative response.

Markers for cell maturation (CD40, CD80, CD86, MHC II) are measured byFACS analysis in the SP37A3 immature mouse dendritic cell (DC) cell lineafter 20 hours of incubation with the test compounds. The assays areperformed as described by Martin et al., Cancer Immuno Immunothe (2014)63(9):925-38, (2014) and Müller et al, Cancer Immunol Res (2014) 2(8),741-55. Compounds are prepared as a 10 mM stock solution in DMSO andsubsequently diluted to the final concentration in cell culture mediumfor use in the cell line studies and were examined using serial dilutionover a concentration range of 1 nM to 10 μM.

Example 3 Induction of In Vitro Cytokine Production by CD4 and CD8 TCells

Five groups including tucaresol, tucaresol plus PD-1 or PD-L1 antibody,tucaresol plus CTLA-4 antibody, CTLA-4 antibody plus PD-1 or PD-L1antibody, and tucaresol plus plinabulin are tested to determine theeffects on in vitro cytokine production by CD4 and. CD8 T cells (e.g,IFN-gamma and IL-2 cells).

The release of pro-inflammatory cytokines (IL-1β, IL-6, IL 12p40) isquantified by ELISA. The assays are performed as described by Martin etal., Cancer Immuno Immunothe (2014) 63(9):925-38. (2014) and Müller etal., Cancer Immunol Res (2014) 2(8), 741-55. Compounds are prepared as a10 mM stock solution in DMSO and subsequently diluted to the finalconcentration in cell culture medium for use in the cell line studiesand are examined using serial dilution over a concentration range of 1nM to 10 μM.

Example 4 Synergy of Tucaresol and Immune Checkpoint Inhibitors (PD-1Antibody/PD-L1 Antibody)

The combined treatment with tucaresol and a PD-1 or PD-L1 checkpointinhibitor is tested in comparison with the treatment with tucaresolalone and the treatment with PD-1 antibody or PD-L1 antibody alone. Thetests are performed using seven to ten-week old immune competent micethat are injected subcutaneously with MC-38 tumor cells. Seven testinggroups are prepared, and each group includes 10 mice.

Group 1 is administered with saline; Group 2 is administered with thetucaresol diluent (in the absence of tucaresol); Group 3 is administeredwith tucaresol dissolved in diluent at a concentration of 5 mg/kg; Group4 is administered with tucaresol dissolved in diluent at a concentrationof 10 mg/kg; Group 5 is administered with PD-1 antibody (or PD-L1antibody); Group 6 is administered with a tucaresol 5 mg/kg/PD-1antibody (or PD-L1 antibody) combined treatment; and Group 7 isadministered with a tucaresol 10 mg/kg/PD-1 antibody (or PD-L1 antibody)combined treatment. For the tucaresol/PD-1 antibody (or PD-L1 antibody)combined treatments (Groups 6 and 7), the mice are administered everyother day for 9 treatments with tucaresol (5 or 10 mg/kg) that isdissolved in diluent, followed by administering PD-1 antibody (or PD-L1antibody) one hour after each tucaresol administration on Days 1 and 3of each week. For the tucaresol only treatment (Groups 3 and 4) or theantibody only treatment (Group 5), mice are administered tucaresol (5 or10 mg/kg dissolved in diluent) every other day for 9 treatments orantibody alone twice per week (Day 1 and Day 3 of each week). For Groups1 and 2, the mice are administered with saline or the tucaresol diluentalone twice per week.

Each treatment starts at tumor size between 10-500 mm³ and continuesuntil Day 24-56. To determine the efficacy of each treatment, thefollowing data are collected mortality rate; the body weight of the miceassessed twice weekly both prior to treatments; the rate of tumor growthas determined by the tumor size measurement (twice every week); thetumor growth index; overall survival rate; the time required to doubletumor size and the tumor weight at necropsy.

Example 5 Synergy of Tucaresol and Immune Checkpoint Inhibitors (PD-1Antibody/PD-L1 Antibody and CTLA-4 Antibody)

The combined treatment with tucaresol, a PD-1 checkpoint inhibitor (orPD-L1 antibody), and a CTLA-4 checkpoint inhibitor is tested incomparison with the treatment with tucaresol alone, the treatment withPD-1 antibody (or PD-Ll antibody) alone, or the treatment with PD-1antibody (or PD-L1 antibody) in combination with CTLA-4 antibody. Thetests are performed using seven to ten-week old immune competent micethat were injected subcutaneously with MC-38 tumor cells. Six testinggroups are prepared, and each group includes 10 mice.

Group 1 is administered with ligEi2a and tucaresol vehicle; Group 2 isadministered with tucaresol dissolved in diluent at a concentration of 5mg/kg; Group 3 is administered with tucaresol dissolved in diluent at aconcentration of 10 mg/kg; Group 4 is administered with PD-1 antibody(or PD-L1 antibody); Group 5 is administered with a tucaresol (5mg/kg)/PD-1 antibody (or PD-L1 antibody) combined treatment; group 6 isadministered with a tucaresol (10 mg/kg)/PD-1 antibody (or PD-LIantibody) combined treatment; Group 7 is administered combined PD-1 (orPD-L1)/CTLA-4 antibodies; and Group 8 is administered with a tucaresol(5 mg/kg)/PD-1 antibody (or PD-L1 antibody)/CTLA-4 antibody combinedtreatment; and Group 9 is administered with tucaresol (10 mg/kg)/PD-1antibody(or PD-L1 andbody)/CTLA-4 antibody combined treatment. For thetucaresol/PD-1 antibody (or PD-L1 antibody) combined treatment (Groups 5and 6) and the tucaresol/PD-1 antibody(or PD-L1 antibody)/CTLA-4antibody treatment (Groups 8 and 9), the mice are administered everyother day with tucaresol (5 or 10 mg/kg) that is dissolved in diluent,for 9 treatments, followed by administering antibody (ies) one hourafter each tucaresol administration on Days 1 and 3 of each week. Forthe tucaresol only treatment (Groups 2 and 3) or the antibody (ies) onlytreatment (Groups 4 and 7), mice are administered with tucaresol (5 or10 mg/kg dissolved in diluent) every other day for 9 treatments orantibody (ies) alone on Day 1 and Day 3 of each week.

Each treatment starts at tumor size between 40-150 mm³ and continuesuntil Day 24-56, when the animals are necropsied. To determine theefficacy of each treatment, the following data are collected: mortality;the body weight of the mice assessed twice weekly both prior totreatments; the rate of tumor growth as determined by the tumor sizemeasurement (twice every week); the tumor growth index; overall survivalrate; the tumor weight at necropsy; and the time required to increasetumor size 10 fold. At necropsy the tissues are weighed and subjected toFACS analysis.

Example 6 Synergy of Tucaresol and Plinabulin

The combined treatment with tucaresol and Plinabulin is tested incomparison with the treatment with tucaresol alone and Plinabulin alone.The tests are performed using seven to ten-week old immune competentmice that are injected subcutaneously with MC-38 tumor cells. Seventesting groups are prepared, and each group includes 10 mice.

Group 1 is administered with saline; Group 2 is administered with thetucaresol diluent (in the absence of tucaresol); Group 3 is administeredwith tucaresol dissolved in diluent at a concentration of 5 mg/kg; Group4 is administered with tucaresol dissolved in diluent at a concentrationof 10 mg/kg; Group 5 is administered with Plinabulin; Group 6 isadministered with a tucaresol 5 mg/kg, and Plinabulin; and Group 7 isadministered with a tucaresol 10 mg/kg and Plinabulin.

Each treatment starts at tumor size between 40-150 mm³ and continuesuntil Day 24-56. To determine the efficacy of each treatment, thefollowing data are collected: mortality rate; the body weight of themice assessed twice weekly both prior to treatments; the rate of tumorgrowth as determined by the tumor size measurement (twice every week);the tumor growth index; overall survival rate; the time required todouble tumor size and the tumor weight at necropsy.

Example 7 Effects in Animal Xenograft Models

Five groups including tucaresol, tucaresol plus PD-1 or PD-L1 antibody,tucaresol plus CTLA-4 antibody, CTLA-4 antibody plus PD-1 or PD-L1antibody, and tucaresol plus plinabulin are tested to determine theireffect in an animal xenograft model.

The combined treatment with tucaresol and the checkpoint inhibitor(s) istested in comparison with the treatment with tucaresol alone, thetreatment with checkpoint inhibitor alone, or combination of checkpointinhibitors. The tests are performed using seven to ten-week old athymic(nu/nu) mice that were injected subcutaneously with human tumor celllines (of either solid or liquid tumor origin, for example of breast,lung, colon, brain, liver, leukemia, myeloma, lymphoma, sarcoma,pancreatic or renal origin). Six to ten testing groups are prepared, andeach group includes 10 mice.

Each treatment starts at tumor size between 40-150 mm³ and continuesuntil Day 24-56, when the animals are necropsied. To determine theefficacy of each treatment, the following data are collected: mortality;the body weight of the mice assessed twice weekly both prior totreatments; the rate of tumor growth as determined by the tumor sizemeasurement (twice every week); the tumor growth index; overall survivalrate; the tumor weight at necropsy; and the time required to increasetumor size 10 fold.

Example 8 Synergy of Tucaresol and Immune Checkpoint Inhibitors

The treatment with the combination of tucaresol and a PD-1 checkpointinhibitor and the combination of tucaresol, a PD-1 antibody, and CTLA-4antibody were tested in comparison with treatment using PD-1 antibodyalone. The tests were performed using seven to ten-week old immunecompetent mice that were injected subcutaneously with MC-38 tumor cells.

Four testing groups were prepared. Group 1 was administered with PD-1antibody (3 mg/kg) alone; Group 2 was administered with a tucaresol (10mg/kg)/PD-1 antibody (3 mg/kg) combined treatment; Group 3 wasadministered with a PD-1 antibody (3 mg/kg)/CTLA-4 antibody (3 mg/kg)combined treatment; Group 4 was administered with a tucaresol (10mg/kg)/PD-1 antibody (3 mg/kg)/CTLA-4 antibody (3 mg/kg) combinedtreatment. For the tucaresol/PD-1 antibody and the tucaresol (10mg/kg)/PD-1 antibody (3 mg/kg)/CTLA-4 antibody (3 mg/kg) combinedtreatments (Groups 2 and 4), the mice were administered tucaresoldissolved in diluent every other day for 9 treatments (10 mg/kg)followed by administering PD-1 antibody one hour after each tucaresoladministration on Days 1 and 3 of each week. For the antibody onlytreatment (Group 1), the mice were administered PD-1 antibody (3 mg/kgdissolved in diluent) twice per week (Day 1 and Day 3 of each week).

Each treatment started at tumor size between 40-150 mm³ and continueduntil Day 24. To determine the efficacy of each treatment, the followingdata were collected: mortality rate; the body weight of the miceassessed twice weekly both prior to treatments; the rate of tumor growthas determined by the tumor size measurement (twice every week); thetumor growth index; overall survival rate; the time required to doubletumor size and the tumor weight at necropsy.

FIG. 1 shows the MC 38 tumor growth in each of the four treatmentgroups. FIG. 2 shows the effect of the four treatment groups on thetumor growth inhibition. FIG. 3 shows the effect of tucaresol incombination with the PD-1 antibody on MC38 tumor weight at necropsy: Asillustrated in these figures, the combination of tucaresol and PD-1antibody had an anti-tumor effect that was similar as or better than thecombination of PD-1 antibody and CTLA-4 antibody, and both combinationsshowed better tumor growth inhibition than PD-1 antibody alone.

Example 9 Comparison of Tucaresol and CTLA-4 Antibody

The treatment with the combination of tucaresol, a PD-1 antibody, andCTLA-4 antibody was compared with the combination of PD-1 antibody andCTLA-4 antibody for their tumor inhibition effects. The tests wereperformed using seven to ten-week old immune competent mice that wereinjected subcutaneously with MC-38 tumor cells.

Three testing groups were prepared. Group A was administered with PD-1antibody (3 mg/kg)/CTLA-4 antibody (3 mg/kg) combined treatment; Group Bwas administered with a tucaresol (10 mg/kg)/PD-1 antibody (3 mg/kg)CTLA-4 antibody (3 mg/kg) combined treatment; and Group C wasadministered with a PD-1 antibody (3 mg/kg)/CTLA-4 antibody (10 mg/kg)combined treatment. For the tucaresol (10 mg/kg)/PD-1 antibody (3mg/kg)/CTLA-4 antibody (3 mg/kg) combined treatments, the mice wereadministered tucaresol dissolved in diluent (10 mg/kg) every other dayfor 9 treatments followed by administering PD-1 antibody one hour aftereach tucaresol administration on Days 1 and 3 of each week. For the PD-1antibody and CTLA-4 antibody group, mice were administered with theantibody twice per week (Day 1 and Day 3 of each week).

Each treatment started at tumor size between 40-150 mm³ and continueduntil Day 24. To determine the efficacy of each treatment, the followingdata were collected: mortality rate; the body weight of the miceassessed twice weekly both prior to treatments; the rate of tumor growthas determined by the tumor size measurement (twice every week); thetumor growth index; overall survival rate; the time required to doubletumor size and the tumor weight at necropsy.

FIG. 4 shows the MC 38 tumor growth in each of the three treatmentgroups. As shown in FIG. 4, when tucaresol (10 mg/kg) was added to acombination of PD-1 (3 mg/kg) and CTLA-4 antibodies, it increased theanti-tumor effect of the combination to a greater extent than increasingthe dose of the CTLA-4 antibody (from 3 mg/kg to 10 mg/kg) in the PD-1antibody and CTLA-4 antibody conibination. The results indicated thattucaresol was more effective in inhibiting tumor growth than CTLA-4antibody when used together with the immune checkpoint inhibitors suchas PD-1 antibody. Because tucaresol has better toxicity and safetyprofile than the CTLA-4 antibody, it can be used as a replacement orsupplement of CTLA-4 antibody in the chemotherapy. The study resultsindicated that adding tucaresol to PD-1 antibody and CTLA-4 antibody canbe more effective against the growth of MC38 tumors than increasing thedose of the CTLA-4 antibody.

What is claimed is:
 1. A pharmaceutical composition, comprising acompound of Formula (I):

Y¹ is selected from hydroxyl, C₁₋₄ alkylamino and acylamino having aC₁₋₄ alkyl moiety thereof; Y², Y³ and Y⁴ are independently selected fromhydrogen, halogen, C₁₋₄ alkyl, C₁₋₄ alkoxy, trifluoromethyl, hydroxyland benzyloxy; and Q¹ is either

where Q² and Q³ are independently selected from hydrogen and C₁₋₄ alkyl;X is selected from cyano, carboxyl or a derivative thereof, 5-tetrazolyland alkylsulfonylcarbamyl having a C₁₋₆ alkyl moiety thereof; and n is 0or an integer selected from 1, 2, 3, 4, 5 and 6, and a pharmaceuticallyacceptable salt thereof; and one or more immune checkpoint inhibitor. 2.The composition of claim 1, wherein the immune checkpoint inhibitor isan inhibitor of PD-1, PD-L1, PD-L2, PD-L3, PD-L4, CTLA-4, LAG3, B7-H3,B7-H4, KIR or TIM3.
 3. The composition of claim 2, wherein the immunecheckpoint inhibitor is a PD-1 inhibitor.
 4. The composition of claim 2,wherein the immune checkpoint inhibitor is a PD-L1 inhibitor.
 5. Thecomposition of claim 2, wherein the immune checkpoint inhibitor is aPD-L2 inhibitor.
 6. The composition of claim 2, wherein the immunecheckpoint inhibitor is a CTLA-4 inhibitor.
 7. The composition of claim1, comprising a first immune checkpoint inhibitor and a second immunecheckpoint inhibitor, wherein the first immune checkpoint inhibitor isdifferent from the second immune checkpoint inhibitor.
 8. Thecomposition of claim 7, wherein the first and the second immunecheckpoint inhibitor is independently an inhibitor of PD-1, PD-L1,PD-L2, PD-L3, PD-L4, CTLA-4, LAG3, B7-H3, B7-H4,KIR or TIM3.
 9. Thecomposition of claim 8, wherein the first immune checkpoint inhibitor isa PD-1 inhibitor, and the second immune checkpoint inhibitor is a CTLA-4inhibitor.
 10. The composition of claim 8, wherein the first immunecheckpoint inhibitor is a PD-L1 inhibitor, and the second immunecheckpoint inhibitor is a CTLA-4 inhibitor.
 11. The composition of claim8, wherein the first immune checkpoint inhibitor is a PD-L2 inhibitor,and the second immune checkpoint inhibitor is a CTLA-4 inhibitor. 12.The composition of any one of claims 1 to 11, wherein the immunecheckpoint inhibitor is an antibody.
 13. The composition of claim 12,wherein the immune checkpoint inhibitor is a PD-1 antibody.
 14. Thecomposition of claim 2, wherein the immune checkpoint inhibitor is aPD-L1 antibody.
 15. The composition of claim 12, wherein the immunecheckpoint inhibitor is a PD-L2 antibody.
 16. The composition of claim12, wherein the immune checkpoint inhibitor is a CTLA-4 antibody. 17.The composition of claim 12, wherein the antibody is selected fromα-CD3-APC, α-CD3-APC-H7, α-CD4-ECD, α-CD4-PB, α-CD8-PE-Cy7,α-CD-8-PerCP-Cy5.5, α-CD11c-APC, α-CD11b-PE-Cy7, α-CD11b-AF700,α-CD14-FITC, α-CD16-PB, α-CD19-AF780, α-CD19-AF700, α-CD20-PO,α-CD25-PE-Cy7, α-CD40-APC, α-CD45-Biotin, Streptavidin-BV605,α-CD62L-ECD, α-CD69-APC-Cy7, α-CD80-FITC, α-CD83-Biotin,Streptavidin-PE-Cy7, α-CD86-PE-Cy7, α-CD86-PE, α-CD123-PE, α-CD154-PE,α-CD161-PE, α-CTLA4-PE-Cy7, α-FoxP3-AF488 (clone 259D),IgG1-isotype-AF488, α-ICOS (CD278)-PE, α-HLA-A2-PE, α-HLA-DR-PB,α-HLA-DR-PerCPCy5.5, α-PD1-APC, VISTA, co-stimulatory molecule OX40, andCD137.
 18. The composition of any one of claims 1 to 17, furthercomprising one or more additional chemotherapeutic agent.
 19. Thecomposition of any one of claims 1 to 18, further comprising one or morepharmaceutically acceptable excipients.
 20. The composition of any oneof claims 1 to 19, further comprising plinabulin.
 21. A pharmaceuticalcomposition, comprising of a compound of formula (I):

Y¹ is selected from hydroxyl, C₁₋₄ alkylamino and acylamino having aC₁₋₄ alkyl moiety thereof; Y², Y³ and Y⁴ are independently selected fromhydrogen, halogen, C₁₋₄ alkyl, C₁₋₄ alkoxy, trifluoromethyl, hydroxyland benzyloxy; and Q¹ is either

where Q² and Q³ are independently selected from hydrogen and C₁₋₄ alkyl;X is selected from cyano, carboxyl or a derivative thereof, 5-tetrazolyland alkylsulfonylcarbamyl having a C₁₋₆ alkyl moiety thereof; and n is 0or an integer selected from 1, 2, 3, 4, 5 and 6, and a pharmaceuticallyacceptable salt thereof; and plinabulin.
 22. The composition of any oneof claims 1 to 21, wherein the compound of Formula (I) is tucaresol. 23.A method for treating cancer, comprising administering thepharmaceutical composition of any one of claims 1 to 22 to a subject inneed thereof.
 24. A method for treating a cancer, comprisingco-administering a compound of Formula (I):

Y¹ is selected from hydroxyl, C₁₋₄ alkylamino and acylamino having aC₁₋₄ alkyl moiety thereof; Y², Y³ and Y⁴ are independently selected fromhydrogen, halogen, C₁₋₄ alkyl, C₁₋₄alkoxy, trifluoromethyl, hydroxyl andbenzyloxy; and Q¹ is either

where Q² and Q³ are independently selected from hydrogen and C₁₋₄ alkyl;X is selected from cyano, carboxyl or a derivative thereof, 5-tetrazolyland alkylsulfonylcarbamyl having a C₁₋₆ alkyl moiety thereof; and n is 0or an integer selected from 1, 2, 3, 4, 5 and 6, and a pharmaceuticallyacceptable salt thereof; and one or more immune checkpoint inhibitor toa subject in need thereof.
 25. The method of claim 24, furthercomprising co-administering one or more additional chemotherapeuticagent.
 26. The method of any one of claims 23 to 25, wherein the cancercomprises cancer cells expressing a binding ligand of PD-1.
 27. Themethod of claim 26, wherein the binding ligand of PD-1 is PD-L1 orPD-L2.
 28. The method of any one of claims 23 to 27, furthercomprisingidentifying cancer cells expressing a binding ligand of PD-1.
 29. Themethod of any one of claims 23 to 27, further comprising identifyingcancer cells expressing PD-L1 or PD-L2.
 30. The method of claim 26,wherein the cancer is head and neck cancer, lung cancer, stomach cancer,colon cancer, pancreatic cancer, prostate cancer, breast cancer, kidneycancer, bladder cancer, ovary cancer, cervical cancer, melanoma,glioblastoma, myeloma, lymphoma, or leukemia.
 31. The method of claim26, wherein the cancer is renal cell carcinoma, malignant melanoma,non-small cell lung cancer (NSCLC), ovarian cancer, Hodgkin's lymphomaor squamous cell carcinoma.
 32. The method of any one of claims 23 to31, wherein the cancer comprises cancer cells expressing a bindingligand of CTLA-4.
 33. The method of claim 32, wherein the binding ligandof CTLA-4 is B7.1 or B7.2.
 34. The method of claim 23 to 33, furthercomprising identifying cancer cells expressing a binding ligand ofCTLA-4.
 35. The method of claim 23 to 33, further comprising identifyingcancer cells expressing B7.1 or B7.2.
 36. The method of any one ofclaims 23 to 35, wherein the immune checkpoint inhibitor is an inhibitorof PD-1, PD-L1, PD-L2, PD-L3, PD-L4, CTLA-4, LAG3, B7-H3, B7-H4, KIR orTIM3.
 37. The method of claim 36, wherein the immune checkpointinhibitor is a PD-1 inhibitor.
 38. The method of claim 36, wherein theimmune checkpoint inhibitor is a PD-L1 inhibitor.
 39. The method ofclaim 36, wherein the immune checkpoint inhibitor is a PD-L2 inhibitor.40. The method of claim 36, wherein the immune checkpoint inhibitor is aCTLA-4 inhibitor.
 41. The method of any one of claims 23 to 40, whereinthe immune checkpoint inhibitor is nivolumab, pembrolizumab,pidilizumab, ipilimumab, dacarbazine, BMS 936559, atezolizumab,durvalimumab, or any combinations thereof.
 42. A method for treatingcancer, comprising co-administering a compound of Formula (I):

Y¹ is selected from hydroxyl, C₁₋₄ alkylamino and acylamino having aC₁₋₄ alkyl moiety thereof; Y², Y³ and Y⁴ are independently selected fromhydrogen, halogen, C₁₋₄ alkyl, C₁₋₄ alkoxy, trifluoromethyl, hydroxyland benzyloxy; and Q¹ is either

where Q² and Q³ are independently selected from hydrogen and C₁₋₄ alkyl;X is selected from cyano, carboxyl or a derivative thereof, 5-tetrazolyland alkylsulfonylcarbamyl having a C₁₋₆ alkyl moiety thereof; and n is 0or an integer selected from 1, 2, 3, 4, 5 and 6, and a pharmaceuticallyacceptable salt thereof; one or more immune checkpoint inhibitor; andplinabulin to a subject in need thereof.
 43. A method for treatingcancer, comprising co-administering a compound of Formula (I):

Y¹ is selected from hydroxyl, C₁₋₄ alkylamino and acylamino having aC₁₋₄ alkyl moiety thereof; Y², Y³ and Y⁴ are independently selected fromhydrogen, halogen, C₁₋₄ alkyl, C₁₋₄ alkoxy, trifluoromethyl, hydroxyland benzyloxy; and Q¹ is either

where Q² and Q³ are independently selected from hydrogen and C₁₋₄ alkyl;X is selected from cyano, carboxyl or a derivative thereof, 5-tetrazolyland alkylsulfonylcarbamyl having a C₁₋₆ alkyl moiety thereof; and n is 0or an integer selected from 1, 2, 3, 4, 5 and 6, and a pharmaceuticallyacceptable salt thereof; and plinabulin to a subject in need thereof.44. The method of claim 42 or 43, wherein the cancer is selected frombreast cancer, colon cancer, rectal cancer, lung cancer, prostatecancer, melanoma, leukemia, ovarian cancer, gastric cancer, renal cellcarcinoma, liver cancer, pancreatic cancer, lymphomas and myeloma. 45.The method of any one of claims 42 to 44, wherein the cancer is a solidtumor or hematological cancer.
 46. The method of any one of claims 42 to45, wherein the cancer does not have any cells expressing PD-1, PD-L1,or PD-L2.
 47. A method of disrupting cancer associated tumor vasculaturein a subject, comprising co-administering a compound of Formula (I):

Y¹ is selected from hydroxyl, C₁₋₄ alkylamino and acylamino having aC₁₋₄ alkyl moiety thereof; Y², Y³ and Y⁴ are independently selected fromhydrogen, halogen, C₁₋₄ alkyl, C₁₋₄ alkoxy, trifluoromethyl, hydroxyland benzyloxy; and Q¹ is either

where Q² and Q³ are independently selected from hydrogen and C₁₋₄ alkyl;X is selected from cyano, carboxyl or a derivative thereof, 5-tetrazolyland alkylsulfonylcarbamyl having a C₁₋₆ alkyl moiety thereof; and n is 0or an integer selected from 1, 2, 3, 4, 5 and 6, and a pharmaceuticallyacceptable salt thereof; and plinabulin.
 48. The method of claim 47,further comprising administering one or more immune checkpointinhibitor.
 49. The method of claim 47 or 48, wherein the cancer isselected from the group consisting of a melanoma, a pancreatic cancer, acolorectal adenocarcinoma, a brain tumor, acute lymphoblastic leukemia,chronic lymphocytic leukemia, hormone refractory metastatic prostatecancer, metastatic breast cancer, non-small cell lung cancer, renal cellcarcinoma, head and neck cancer, prostate cancer, colon cancer,anaplastic thyroid cancer,
 50. The method of any one of claims 23 to 49,wherein the compound of Formula (I) is tucaresol.